Ultrasound delivery of nanoparticles

ABSTRACT

Enhanced delivery of compositions for treatment of skin tissue with photoactive plasmonic nanoparticles and light, with embodiments relating to delivery devices using, for example, ultrasound. Treatments are useful for cosmetic, diagnostic and therapeutic applications.

INCORPORATION BY REFERENCE

This application is a continuation of U.S. patent application Ser. No.14/321,509 filed Jul. 1, 2014, now U.S. Pat. No. 9,572,880, which is acontinuation in part of U.S. patent application Ser. No. 14/020,423filed Sep. 6, 2013, now U.S. Pat. No. 8,834,933, which is a continuationof U.S. patent application Ser. No. 13/219,514 filed Aug. 26, 2011, nowU.S. Pat. No. 9,061,056, which claims the benefit of priority of U.S.Provisional Application Nos. 61/402,305 filed Aug. 27, 2010; 61/422,612filed Dec. 13, 2010, and 61/516,308 filed Apr. 1, 2011; each of which ishereby incorporated by reference in its entirety. This application alsoclaims the benefit of priority of U.S. Provisional Application No.61/870,103 filed Aug. 26, 2013. Any and all applications for which aforeign or domestic priority claim is identified in the Application DataSheet as filed with the present application are hereby incorporated byreference under 37 CFR 1.57.

PARTIES OF JOINT RESEARCH AGREEMENT

The invention described herein was created subject to a Joint ResearchAgreement between Sienna Labs, Inc. and Nanocomposix, Inc.

BACKGROUND

Field of the Invention

The field of the invention comprises nanoparticles and/or photoactivecompounds for use in cosmetic, diagnostic and/or therapeutic procedures,including ultrasonic delivery systems and methods for delivering theparticles and/or compounds to a target tissue. Ultrasound, according toseveral embodiments, is low frequency ultrasound delivered by, forexample, a transducer or sonotrode. In several embodiments, theinvention relates to using laser or light energy combined withnanoparticles and/or photoactive compounds to modify, smooth, and/orresurface the skin (including tissue under the skin surface) of humans.

Description of the Related Art

Laser treatments of the skin have been highly touted for therapeutic andcosmetic utility. Therapeutically, potential uses for laser skin therapyinclude laser ablation of cancerous cells in cancer patients and laserablation of damaged tissue in burn victims. Cosmetic applications forlaser skin therapy are much more numerous, and include hairremoval/reduction, treatment of dyschromia, shrinking of the skinfollowing operations such as liposuction, acne treatment, chemical orphysical abrasion of unwanted markings on the skin, surgical treatmentsincluding nose reduction and face- and neck-lifts, and other aestheticskin remodeling purposes.

SUMMARY

In several embodiments, the invention relates to using laser or lightenergy combined with nanoparticles and/or photoactive compounds to treatthe skin (including tissue under the skin surface) using ultrasound tofacilitate the delivery of the nanoparticles and/or photoactivecompounds. In some embodiments, the invention is able to modify, smooth,and/or resurface the skin (including tissue under the skin surface) ofhumans. Several embodiments of the present invention also relate tomethods for focusing electromagnetic energy with particles and/orphotoactive compounds to selectively heat target regions of skin with adiscrete, often minute, size and shape for the treatment of acne,especially acne scars, while not damaging surrounding skin tissue. Insome embodiments, the particles can be microparticles and/ornanoparticles.

Also, provided herein, in several embodiments, are compositions andmethods useful in the targeted thermomodulation of target cellpopulations and target tissues, for the purposes of cosmetic treatmentsand the treatment and prevention of chronic and acute diseases anddisorders and enhanced delivery systems and methods (e.g., usingultrasound) for delivering the particles and/or compounds to a targettissue.

In various embodiments, a method of delivering a composition to a targettissue under a skin surface with a delivery device includes applying acomposition to a skin surface, and distributing the composition from theskin surface to a target tissue under the skin surface with a deliverydevice. In one embodiment, the delivery device is an ultrasound device.In one embodiment, the composition comprises a plurality of unassembledplasmonic nanoparticles. In one embodiment, the unassembled plasmonicnanoparticles comprise a conductive metal portion. In one embodiment,the conductive metal portion comprises at least one of gold or silver.In one embodiment, the unassembled plasmonic nanoparticles have a sizein a range of 10 nm to 300 nm. In one embodiment, the unassembledplasmonic nanoparticles comprise a coating that coats the conductivemetal portion, wherein the coating facilitates selective removal fromthe skin surface. In one embodiment, the coating comprises at least oneof silica or polyethylene glycol (PEG). In one embodiment, theunassembled plasmonic nanoparticles have a concentration of 10⁹ to 10²³particles per ml of the composition, wherein the concentration issufficient to, after exposure to irradiation, induce thermal damage in asebaceous gland. In one embodiment the method includes selectivelyremoving the composition from the skin surface, while leaving thecomposition localized within the sebaceous gland. In one embodiment themethod includes irradiating the composition with an infrared lightsource thereby inducing a plurality of surface plasmons in theunassembled plasmonic nanoparticles. In one embodiment, the plurality ofsurface plasmons generates localized heat in the target tissue. In oneembodiment, the mechanical vibration device is a low frequencyultrasound device. In one embodiment, the method includes pre-treatingthe skin surface prior to irradiating the composition, whereinpre-treating the skin surface comprises hair removal. In one embodiment,the unassembled plasmonic nanoparticles comprise an optical density of10 O.D. to 5,000 O.D. at an infrared light range. In one embodiment, theunassembled plasmonic nanoparticles comprise a solid, conducting silvercore and a silica coating. In one embodiment, the conductive metalportion is a silver nanoplate, and the coating is less conductive thanthe conductive metal portion. In one embodiment, the conductive metalportion is a nanoplate, and the nanoplate has a peak absorptionwavelength in a range of 750 nm to 1200 nm. In one embodiment, thecoating comprises silica, wherein generation of localized heat issufficient to affect at least one of a sebocyte and sebum.

In various embodiments, a method of delivering a composition to asebaceous gland includes topically applying a solution of unassembledplasmonic nanoparticles to a skin surface, and targeting a sebaceousgland by redistributing the solution of unassembled plasmonicnanoparticles from the skin surface to the sebaceous gland with adelivery device.

In one embodiment, the delivery device is a mechanical vibration device,wherein the mechanical vibration device comprises at least one of thegroup consisting of an ultrasound device and a massage device. In oneembodiment, the unassembled plasmonic nanoparticles have a dimension ina range of 10 nm to 300 nm. In one embodiment, the unassembled plasmonicnanoparticles have a concentration of 10⁹ to 10²³ particles per ml ofthe solution. In one embodiment, the unassembled plasmonic nanoparticlescomprise a conductive metal portion. In one embodiment, the conductivemetal portion comprises at least one of gold or silver. In oneembodiment, the unassembled plasmonic nanoparticles comprise a coatingthat coats the conductive metal portion, wherein the coating facilitatesselective removal from the skin surface. In one embodiment, the coatingcomprises at least one of silica or polyethylene glycol (PEG). In oneembodiment, the method includes selectively removing the solution fromthe skin surface, while leaving the solution localized within thesebaceous gland. In one embodiment, the method includes irradiating thesolution of unassembled plasmonic nanoparticles with an energywavelength in a range of 750 nm to 1200 nm to induce a plurality ofsurface plasmons in the unassembled plasmonic nanoparticles, therebytreating acne at the sebaceous gland. In one embodiment, the mechanicalvibration device is configured for bubble formation or liquidmicrostreaming. In one embodiment, the method includes pre-treating theskin surface to increase delivery of the unassembled plasmonicnanoparticles to the sebaceous gland with at least one of the groupconsisting of shaving, waxing, peeling, cyanoacrylate surface peeling, acalcium thioglycolate treatment, a surface exfoliation, a mechanicalexfoliation, a salt glow, a microdermabrasion, a chemical exfoliation, achemical exfoliation with an enzyme, a chemical exfoliation withalphahydroxy acid, and a chemical exfoliation with betahydroxy acid. Inone embodiment, the concentration of the unassembled plasmonicnanoparticles is 10⁹ to 10¹⁶ particles per ml of the solution. In oneembodiment, the coating is less conductive than the conductive metalportion. In one embodiment, the unassembled plasmonic nanoparticles havean optical density of 10 O.D. to 5,000 O.D. within an infrared lightrange. In one embodiment, the coating is semiconductive, wherein theconductive metal portion is inside the coating, and wherein the coatingis less conductive than the conductive metal portion. In one embodiment,the conductive metal portion is a nanoplate, and wherein the coating isless conductive than the conductive metal portion.

In various embodiments, a method of delivering a composition ofunassembled plasmonic nanoparticles to a pilosebaceous unit includesapplying a solution of unassembled plasmonic nanoparticles to a skinsurface, and distributing the solution of unassembled plasmonicnanoparticles with a mechanical vibration device from the skin surfaceto a pilosebaceous unit thereby targeting the pilosebaceous unit. In oneembodiment, the mechanical vibration device comprises at least one ofthe group consisting of an ultrasound device, a sonic force device, amassage device, a high pressure air flow device, a high pressure liquidflow device, and a vacuum device, and a dermabrasion device. In oneembodiment, the pilosebaceous unit comprises one or more structuresconsisting of: a hair shaft, a hair follicle, a sebaceous gland, and ahair follicle infundibulum. In one embodiment, the unassembled plasmonicnanoparticles comprise a conductive metal portion. In one embodiment,the conductive metal portion comprises at least one of gold or silver.In one embodiment, the unassembled plasmonic nanoparticles have a peakabsorption wavelength of between 750 nm and 1200 nm. In one embodiment,the unassembled plasmonic nanoparticles have a concentration of 10⁹ to10²³ particles per ml of the solution. In one embodiment, theunassembled plasmonic nanoparticles comprise a coating that coats theconductive metal portion. In one embodiment, the coating comprises atleast one of silica or polyethylene glycol (PEG). In one embodiment, themethod includes selectively removing the solution from the skin surfacewhile leaving the solution localized within the portion of the sebaceousgland. In one embodiment, the method includes irradiating the solutionwith an energy to induce the unassembled plasmonic nanoparticles togenerate localized thermal damage in the sebaceous gland. In oneembodiment, the method includes pre-treating the skin surface toincrease delivery of the unassembled plasmonic nanoparticles to thepilosebaceous unit with at least one of the group consisting of shaving,waxing, peeling, cyanoacrylate surface peeling, a calcium thioglycolatetreatment, a surface exfoliation, a mechanical exfoliation, a salt glow,a microdermabrasion, a chemical exfoliation, a chemical exfoliation withan enzyme, a chemical exfoliation with alphahydroxy acid, and a chemicalexfoliation with betahydroxy acid. In one embodiment, the methodincludes irradiating the solution of unassembled plasmonic nanoparticlescomprises exposing the solution of unassembled plasmonic nanoparticlesto the energy at a wavelength of between 750 nm and 1200 nm to induce aplurality of surface plasmons in the unassembled plasmonicnanoparticles, thereby treating acne at the sebaceous gland. In oneembodiment, the mechanical vibration device comprises at least a lowfrequency ultrasound device. In one embodiment, the method includesirradiating the solution of unassembled plasmonic nanoparticles with theenergy comprises an infrared light source wavelength of between 750 nmand 1200 nm to induce a plurality of surface plasmons in the unassembledplasmonic nanoparticles, thereby treating acne at the sebaceous gland.In one embodiment, the unassembled plasmonic nanoparticles arenanoplates. In one embodiment, the unassembled plasmonic nanoparticleshave an optical density of 10 O.D. to 5,000 O.D. within an infraredlight range and the concentration is 10⁹ to 10¹⁸ particles per ml of thesolution. In one embodiment, the method includes irradiating thesolution of unassembled plasmonic nanoparticles with the energycomprises an infrared wavelength of between 750 nm and 1200 nm to inducea plurality of surface plasmons in the unassembled plasmonicnanoparticles, thereby heating the pilosebaceous unit. In oneembodiment, selectively removing the composition from the skin surfacecomprises using water or alcohol to remove the composition from the skinsurface while leaving the composition localized within the pilosebaceousunit.

Several embodiments of the present invention provide safe, tolerable,and efficacious treatments for acne and acne scarring that achieveprolonged improvement of the skin. Other light based treatments foracne, including photodynamic therapy (PDT) and long wave length lasers(e.g. 1450 nm), tend to need high energy illumination and may lacktarget specificity, which can lead to intolerable off-targetside-effects including sensitivity to light, pain, inflammation,hyper/hypo-pigmentation, and permanent scarring. Many traditional lightbased procedures for treating acne scars, including ablative andnon-ablative skin resurfacing, often involve aggressive treatmentsettings that lead to long healing times and risk of side-effect (e.g.,hyperpigmentation, scarring). Several embodiments of the presentinvention are particularly effective for box car scars, ice pick scars,and other pitted scars, where excision is otherwise considered among theonly reliable methods for treatment.

Human skin is vulnerable to damage, scarring, and an overall decline inskin smoothness or texture from disease, trauma, environmental exposure,and aging. Consumer demand for aesthetic skin enhancement that hasminimal risk and provides rapid recovery has resulted in efforts toprovide methods of non-surgical skin rejuvenation including skinresurfacing (e.g., lasabration, laser peel and laser vaporization).However, many skin resurfacing and other techniques resulting in theremoval of epidermal layers fail to address deeper, dermal-layer scarsand skin lesions.

Skin resurfacing generally involves controlled removal and, optionally,regeneration of the skin either from ablative or non-ablative damage. Ingeneral, these and related aesthetic procedures use electro-magneticenergy and/or heat to induce thermal injury in areas of the skin, andare often considered minimally invasive. Much of the prior work in skinresurfacing involves either non-fractional skin resurfacing orfractional skin resurfacing.

Non-fractional skin resurfacing uses non-fractional high-energy pulsedand/or scanned CO₂ or Er:YAG lasers, the energy from which when directedto the skin remove skin material in a controlled manner. Severalembodiments of the present invention are particularly advantageousbecause some or all of the following advantages are present: (i)prolonged and unpleasant postoperative recovery period characterized byedema, oozing, and burning discomfort are avoided, (ii) substantialpatient pain and discomfort during the procedure, generally requiring asignificant amount of analgesia (local anesthetic for nerve blockade orgeneral anesthesia) are not needed; (iii) reduced incidence ofcomplications including persistent erythema, hyperpigmentation,hypopigmentation, scarring, and infection (e.g., infection with Herpessimplex virus); (iv) ability to selectively and uniformly target energyto small target regions, e.g., lesions or scars with dimensions of a fewmm²; (v) reduced incidence of physical impediments (“shadows”) thatprevent uniform delivery of the laser energy to desired areas in thetarget skin (which can occur because energy from certain non-fractionallasers is absorbed by water in the first few tissue layers of skincells, thus causing any irregular contours on the skin surface (e.g.deep ice-pick or box car scars) to form shadows); (vi) reducedappearance of visible spots and/or edges after treatment due toinflammation, pigmentation, or texture changes (e.g., as would otherwisecorrespond to the sites of treatment including the specific edges of thelaser spot); and (vii) reduced risk of irregularity in wound healing,inconsistent skin regeneration and potential scarring (which can occur,for example, when overlapping regions are treated by non-fractionallasers).

A derivative technique from the ablative use of non-fractional lasersfor skin resurfacing is to induce selective thermal damage to thesub-epidermal layer (particularly the dermal layer) with no disruptionof the superficial epidermal layer integrity. Such techniques have beentermed non-ablative resurfacing, non-ablative subsurfacing, ornon-ablative skin remodeling, and have been used as an alternativeprocedure. Techniques generally utilize non-ablative lasers, flashlamps,or radio frequency currents. However, while some of the adverse effectsfrom epidermal ablation may be reduced, overall efficacy is typicallylimited because of epidermal sparing. Several embodiments of theinvention are advantageous because irregular patterns of energydeposition are reduced or absent, thereby reducing or avoiding visiblespots or edges.

Fractional resurfacing has emerged as a technique attempting to addresssome of the limitations in patient discomfort and recovery time fromnon-fractional approaches. In fractional resurfacing, thermally ablatedmicroscopic zones of epidermis and dermis (referred to as “micro thermalzones”) are spaced in a grid over the skin surface in a generallycontrolled, geometric pattern; the non-ablated zones in the uninjuredsurrounding tissue serves as a reservoir of cells that accelerate andpromote safe and rapid healing. The affected zones can compromiseapproximately 15-70% of the skin surface area per treatment session andcan be randomly selected by the orientation of the geometric pattern.Several embodiments of the present invention are particularlyadvantageous because (i) pre-determined arrays are not needed, and thus,all or substantially all of a small target region such as a scar regionthat does not exceed about 25 mm² (e.g., a scar region that is withinabout 1-10 mm², 10-15 mm², 15-25 mm², and ranges therein) can betreated; (ii) skin outside of target regions do not need to beunnecessarily treated and the selective targeting of small regions canbe achieved with respect to scars or other lesions (whether atrophic ornot) where new collagen formation and re-epithelialization would providethe highest benefit; (iii) several embodiments provide a balance betweenaggressive treatments with high skin surface coverage that arecharacterized by long healing time and less aggressive treatments thathave no effect; and (iv) several embodiments apply electromagneticenergy to selectively and uniformly heat small target regions that arenear or below the spot size of the energy (e.g., light), while sparingother tissue.

In general, traditional methods involving light and lasers are promisingfor the treatment skin disorders, but are still insufficientlyeffective. Ultraviolet (UV)/blue light is approved by the FDA for thetreatment of mild to moderate acne only, due to its anti-inflammatoryeffects mediated on skin cells (keratinocytes), potentially through theaction of endogenous porphyrin photosensitizers within follicles.Exogenous porphyrin precursors such as 5-aminoluveulinic acid (5-ALA)have been formulated for topical or oral delivery and shown toaccumulate within sebaceous follicles, absorb photons from red lightexposure and form reactive oxygen species that directly damage cellularmembranes and proteins. This procedure combining porphyrin applicationand high intensity red light, termed ‘photodynamic therapy’, has beendemonstrated to reduce sebum production and acne by 50% for 20 weekspost-irradiation. However, high intensity energies (50-150 J/cm²) arerequired to damage sebaceous gland skin structures, and transdermalporphyrin penetration leads to off-target side-effects which includesensitivity to light, pain, inflammation, hyper/hypo-pigmentation, andpermanent scarring. Several embodiments of the invention areparticularly advantageous because they locally induce photo-destructionin skin structures without affecting surrounding tissues.

Other advantages of several embodiments of the invention include reducedor no procedural pain and discomfort, post-procedural discomfort,lengthy recovery time, post-procedural infection, and unintendedscarring. Additional advantages include reduced or no non-specific skindamage, skin irritation and scarring. Although many advantages aredescribed herein, all of these advantages need not be present in any oneembodiment. Several embodiments of the invention address the need toprovide a procedure that enables patient-specific focusing ofelectromagnetic energy and heat in a manner to effectively damage smalltarget regions while minimizing damage to surrounding tissue, therebyproviding improved efficacy in remodeling target areas whilesubstantially reducing or eliminating undesirable side effects such asprocedural discomfort, post-procedural discomfort, a lengthy healingtime, post-procedural infection, and unintended scarring. Severalembodiments described herein provide the use of concentrated light forskin resurfacing and scar removal with less pain, and that is faster,easier and more efficacious.

Several embodiments of the invention are useful for hair removal and/orreduction. Light-based hair removal systems suffer from particularly lowrates of efficacy at removing light hair (vellus, blonde, gray, redhair). Multiple (even 6 or more) treatments are insufficient to achievea therapeutic result in blonde- gray- or red-haired patients, even withthe use of topically applied chromophores such as carbon. In addition tolight hair removal, thermoablative technology, as described in severalembodiments of the invention herein, has untapped potential in thefields of wound healing, tissue remodeling, vascular repair, and acnetreatment.

As described herein, many embodiments of the invention are used fortreating acne and the scars that result from acne. Acne vulgaris resultsfrom obstruction of the pilosebaceous unit, consisting of the hairshaft, hair follicle, sebaceous gland and erector pili muscle, whichleads to accumulation of sebum oil produced from the sebaceous gland andthe subsequent colonization of bacteria within the follicle.Microcomedones formed as a result of accumulated sebum progress tonon-inflamed skin blemishes (white/blackheads), or to skin blemisheswhich recruit inflammatory cells and lead to the formation of papules,nodules and pus-filled cysts. The sequelae of untreated acne vulgarisoften include hyperpigmentation, scarring and disfiguration, as well assignificant psychological distress. Therefore, acne treatments seekbroadly to reduce the accumulation of sebum and microorganisms withinfollicles and the sebaceous gland. In several embodiments of theinvention, reduction of microorganisms, via the photoactive particles(e.g., plasmonic nanoparticles) described herein, include, but is notlimited to, inactivation of bacteria or other microorganisms, reductionin the number, growth, viability, and/or function etc. of bacteria orother microorganisms. This reduction can be accomplished by, forexample, the heat generated by several of the embodiments describedherein and/or the enhanced delivery of drugs and other substances. Thisreduction can be accomplished in target regions, including atrophicregions and small target regions as described herein.

In one aspect, described herein are compositions of matter. For example,in one embodiment, provided is a composition comprising a cosmeticallyacceptable carrier and a plurality of photoactive particles (e.g.,plasmonic nanoparticles) in an amount effective to inducethermomodulation in a target tissue region with which the composition istopically contacted.

In some embodiments, the composition comprises, or consists essentiallyof photoactive particles (e.g., plasmonic nanoparticles) that areactivated by exposure to energy delivered from a surface plasmonresonance excitation sources (e.g., nonlinear excitation surface plasmonresonance source) to the target tissue region. As discussed herein, atresonance wavelengths plasmonic nanoparticles can act as antennas,providing a “nonlinear excitation” at peak resonance or, in other words,an enhanced extinction cross section for a given physical cross-sectionof material when compared to non-plasmonic photoactive materials of thesame dimension. Thus, in several embodiments, plasmonic materials areable pull more energy from delocalized electromagnetic waves surroundingthe material at peak resonance than non-plasmonic photoactive materialof the same dimension.

In further or additional embodiments, described herein are compositionscomprising, or consisting essentially of, at least one photoactiveparticles (e.g., plasmonic nanoparticle) that comprises a metal,metallic composite, metal oxide, metallic salt, electric conductor,electric superconductor, electric semiconductor, dielectric, quantum dotor composite from a combination thereof. In further or additionalembodiments, provided herein is a composition wherein a substantialamount of the photoactive particles (e.g., plasmonic particles) presentin the composition comprise geometrically-tuned nanostructures. Incertain embodiments, provided herein is a composition whereinphotoactive particles (e.g., plasmonic particles) comprise any geometricshape currently known or to be created that absorb light and generateplasmon resonance at a desired wavelength, including nanoplates, solidnanoshells, hollow nanoshells, partial nanoshells, nanorods, nanorice,nanospheres, nanofibers, nanowires, nanopyramids, nanoprisms, nanostars,nanocrescents, nanorings, or a combination thereof. In yet additionalembodiments, described herein is a composition wherein the photoactiveparticles (e.g., plasmonic particles) comprise silver, gold, nickel,copper, titanium, silicon, galadium, palladium, platinum, or chromium,as well as including metal alloys, composites, and amalgams.

In some embodiments, provided herein is a composition comprising acosmetically acceptable carrier that comprises, or consists essentiallyof, an additive, a colorant, an emulsifier, a fragrance, a humectant, apolymerizable monomer, a stabilizer, a solvent, or a surfactant. In oneembodiment, provided herein is a composition wherein the surfactant isselected from the group consisting of: sodium laureth 2-sulfate, sodiumdodecyl sulfate, ammonium lauryl sulfate, sodium octech-1/deceth-1sulfate, lipids, proteins, peptides or derivatives thereof. In oneembodiment, provided is a composition wherein a surfactant is present inan amount between about 0.1 and about 10.0% weight-to-weight of thecarrier. In yet another embodiment, the solvent is selected from thegroup consisting of water, propylene glycol, alcohol, hydrocarbon,chloroform, acid, base, acetone, diethyl-ether, dimethyl sulfoxide,dimethylformamide, acetonitrile, tetrahydrofuran, dichloromethane, andethylacetate. In one embodiment, the composition comprises, or consistsessentially of, photoactive particles (e.g., plasmonic particles) thathave an optical density of at least about 1 O.D. at one or more (e.g.,50 O.D.-10,000 O.D.) peak resonance wavelengths at, for example,infrared.

In further or additional embodiments, described herein is a compositionwherein photoactive particles (e.g., plasmonic particles) comprise ahydrophilic or aliphatic coating, wherein the coating does notsubstantially adsorb to skin of a mammalian subject, and wherein thecoating comprises polyethylene glycol, silica, silica-oxide,polyvinylpyrrolidone, polystyrene, polyquaternium(s), a protein or apeptide. In yet an additional embodiment, the thermomodulation comprisesdamage, ablation, thermoablation, lysis, denaturation, deactivation,activation, induction of inflammation, activation of heat shockproteins, perturbation of cell-signaling or disruption to the cellmicroenvironment in the target tissue region. Still further, in certainpresentations the target tissue region comprises a sebaceous gland, acomponent of a sebaceous gland, a sebocyte, a component of a sebocyte,sebum, or hair follicle infundibulum. In further embodiments, the targettissue region comprises a bulge, a bulb, a stem cell, a stem cell niche,a dermal papilla, a cortex, a cuticle, a hair sheath, a medulla, anarrector pili muscle, a Huxley layer, or a Henle layer.

In another aspect, described herein are methods of performing targetedablation of tissue. For example, in one embodiment, provided is a methodfor performing targeted ablation of a tissue to treat a mammaliansubject in need thereof, comprising the steps of i) topicallyadministering to a skin surface of the subject a composition ofphotoactive particles (e.g., plasmonic particles) ii) providingpenetration means to redistribute the plasmonic particles from the skinsurface to a component of dermal tissue; and iii) causing irradiation ofthe skin surface by light. In further or additional embodiments,provided is a method wherein the light source comprises excitation ofmercury, xenon, deuterium, or a metal-halide, phosphorescence,incandescence, luminescence, light emitting diode, or sunlight. In stillfurther or additional embodiments, provided is a method wherein thepenetration means comprises high frequency ultrasound, low frequencyultrasound (e.g., frequencies of 1 kHz to 500 kHz, e.g., 1 kHz-100 kHz,5 kHz-45 kHz, 20 kHz-50 kHz, 30 kHz-40 kHz, 30 kHz, 40 kHz, and anyranges or frequencies therein), massage (e.g., hand massage, vibration,mechanical vibration, and/or at frequencies of less than 1 kHz, 1 Hz-900Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250 Hz, and any frequenciestherein), iontophoresis, high pressure air flow, high pressure liquidflow, vacuum, pre-treatment with fractionated photothermolysis ordermabrasion, or a combination thereof, and/or pre-treatment with heat,massage, ultrasound or a combination thereof. In still furtherembodiments, provided is a method wherein the irradiation compriseslight having a wavelength of light between about 200 nm and about 10,000nm (e.g., 700 nm to 1,200 nm, 600 nm to 1,500 nm, 500 nm to 2,000 nm), afluence of about 0.1 to about 100 joules/cm² (e.g., 1 to 60 joules/cm²,5 to 50 joules/cm² 10 to 30 joules/cm²), a pulse width of about 1femptosecond to about 1 second (e.g., 100 microsecond to 500millisecond, 100 microsecond to 1000 microseconds, 1 millisecond to 10millisecond, 10 millisecond to 100 millisecond, 100 millisecond to 500millisecond), and a repetition frequency of about 1 Hz to about 1 THz(e.g., 1 Hz to 10 Hz, 1 Hz to 1 MHz, 1 Hz to 1 GHz).

In a further aspect, provided herein are some embodiments ofcompositions comprising a cosmetically acceptable carrier, an effectiveamount of sodium dodecyl sulfate, and a plurality of photoactiveparticles (e.g., plasmonic nanoparticles) in an amount effective toinduce thermal damage in a target tissue region with which thecomposition is topically contacted, wherein the nanoparticles have anoptical density of at least about 1 O.D. (e.g., 10 O.D., 50 O.D., 100O.D., 1000 O.D., and/or 10,000 O.D.) at a resonance wavelength in therange of about 810 nanometers or 1064 nanometers, wherein the plasmonicparticles comprise a silica coating from about 5 to about 35 nanometers,wherein the acceptable carrier comprises water and propylene glycol. Inyet another aspect, provided are some embodiments of systems for laserablation of hair or treatment of acne comprising a composition and asource of plasmonic energy suitable for application to the human skin.

In several embodiments, the invention comprises a method for reducingdermal scar tissue (e.g., in a human subject) comprising: i) identifyinga target region of skin tissue on a human subject, wherein the targetregion comprises an epidermal surface and dermal scar tissue, whereinthe target region does not exceed about 25 mm²; (ii) contacting theepidermal surface of the target region with a non-dispersive compositioncomprising a photoactive material; and (iii) delivering to the targetregion energy in the 700 nm to about 1200 nm range in an amountsufficient to heat at least a portion of the dermal scar tissue to atemperature of at least 40 degrees Celsius for a period of timesufficient to reduce the dermal scar tissue. The temperature may be inthe range of about 40 to 45, 40 to 50, 40 to 55, 40 to 60, 40 to 65, 40to 70, 40 to 75, 40 to 80 or 40 to above 80. The time period may be inthe range of 1 femptosecond to about 1 second (e.g., 100 microsecond to1000 microseconds, 1 millisecond to 10 millisecond, 10 millisecond to100 millisecond, 100 millisecond to 500 millisecond).

In some embodiments, the invention comprises a method for reducingdermal scar tissue (e.g., in a human subject) comprising: (i)identifying a target region of skin tissue on a human subject, whereinthe target region comprises an epidermal surface and dermal scar tissuecomprising a pathophysiological collagen deposition, dermal matrix, orepidermal surface, wherein the target region does not exceed about 25mm²; (ii) contacting the epidermal surface of the target region with anon-dispersive composition comprising a photoactive material; and (iii)delivering to the target region energy in the 700 nm to about 1200 nmrange in an amount sufficient to heat at least a portion of the dermalscar tissue to a temperature sufficient to cause damage andregeneration, thereby reducing the dermal scar tissue.

For embodiments in which the target region (e.g., atrophic scar) doesnot exceed about 25 mm² the region may be sized in at least onedimension as follows: 0.1 mm² to 5 mm², 5 mm²-10 mm², 10 mm²-15 mm², 15mm²-25 mm², 0.1 mm²-25 mm² overlapping ranges therein. These dimensionscan apply, for example, to the surface area of the region (the innersurface area of an atrophic scar). The target region (e.g., atrophicscar) may also have at least one dimension (e.g., depth, length, width)in the following ranges: 0.01 mm-10 mm (e.g. 0.5 mm to 1.5 mm, 0.25mm-2.5 mm, 1 mm to 8 mm, 5 mm to 10 mm, and overlapping ranges therein).In one embodiment, the target region (e.g., atrophic scar) is at least0.25 mm mean thickness (e.g., 0.01-0.25 mm).

In several embodiments, the methods can be performed in any order, withany step repeated one or more times. In some embodiments of the methods,the contacting and/or delivering steps can be repeated 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 15, 20, 24, or more times. In several embodiments, themethods for treating the target regions are repeated one or more timeson one or more additional target regions. For example, the procedure maybe performed/repeated 1-24 times (e.g., 2, 3, 4, 5, 10 or more times). Asingle target region may be treated, or alternatively, multiple targetregions may be treated sequentially or simultaneously.

In several embodiments, the dermal scar tissue comprises a scarresulting from an acne vulgaris infection. The dermal scar tissue may beatrophic (e.g., recessed). The target region of skin tissue is locatedon the face or neck of a human subject.

In several embodiments, the photoactive material comprises carbon. Inseveral embodiments, the photoactive material comprises graphite. Inseveral embodiments, the photoactive material comprises a plasmonicnanoparticle. In several embodiments, the photoactive material comprisesa silver plasmonic nanoparticle. In several embodiments, the photoactivematerial comprises a silica-coated silver plasmonic nanoparticle. Inseveral embodiments, the photoactive material is present at aconcentration of from about 0.01% to about 10% volume to volume ratio,or greater than 10% volume to volume ratio (e.g., 0.01%-0.1%, 0.1%-1%,1%-10%. In several embodiments, the photoactive material does notsubstantially penetrate the epidermal surface. In several embodiments,the energy comprises a spot diameter anywhere in the range of about 0.5mm to about 20 mm at the epidermal surface.

In several embodiments, the non-dispersive composition comprises avolume of about 10 microliters with a diameter of less than about 5 mmat the epidermal surface for at least one minute after contacting of thenon-dispersive composition with the epidermal surface. In severalembodiments, the non-dispersive composition does not laterally migratealong the epidermal surface at a rate greater than about 1 mm perminute. In several embodiments, the non-dispersive composition comprisesat least one of water, a humectant, a surfactant, a thickener, a dye, anantiseptic, an anti-inflammatory agent, an anti-oxidant, a vitamin, afragrance, an oil, or a topical anesthetic. In several embodiments, thenon-dispersive composition is contacted with the epidermal surface witha volume of about 1 to about 50 microliters of the non-dispersivecomposition.

In several embodiments, the methods further comprise the step ofcontacting the epidermal surface of the target region with an adhesivecompound prior to contacting the epidermal surface with thenon-dispersive composition, wherein the adhesive compound increasesretention of the photoactive material at the target region.

In several embodiments, the invention comprises a means for deliveringphotoactive particles into small target regions. In several embodiments,the means for delivering includes an apparatus for delivering aformulation into a target region (e.g., an acne scar or other atrophicscar) of a human subject. As used herein, the terms formulation andcomposition can be used interchangeably. In several embodiments, theapparatus, comprises a supply of a liquid formulation comprising anphotoactive material, which, when put in substantial physical contactwith a target area of a skin surface of a human subject, with the targetarea comprising an acne scar or portion thereof, is capable ofpenetrating the skin surface at the target area to denature at least onepathophysiological collagen deposition present in the acne scar, bydelivering sufficient thermal energy to the targeted area such that thetemperature of the collagen deposition in the target area is elevatedabove the denaturation temperature of the collagen deposition. Inseveral embodiments, the apparatus enables the liquid formulation, whenthe liquid formulation is put in contact with the target area, to besubstantially retained in the target area, wherein the apparatusdelivers the liquid formulation onto the target area in a volume of fromabout 0.01 ml to about 1 ml.

In several embodiments, the invention comprises a system for affectingcollagen (e.g., denaturing collagen) present in a target region such asan atrophic region (e.g., acne scar) comprising: (i) an apparatus fordelivering a formulation, and (ii) a light source. In severalembodiments, the apparatus contains a supply of a liquid formulationcomprising an photoactive material, which, when put in substantialphysical contact with a target area of a skin surface of a humansubject, the target area comprising an acne scar or portion thereof, iscapable of penetrating the skin surface at the target area to denatureat least one pathophysiological collagen deposition present in the acnescar, by delivering sufficient thermal energy to the targeted area suchthat the temperature of the collagen deposition in the target area iselevated above the denaturation temperature of the collagen deposition.In several embodiments the apparatus enables the liquid formulation,when the liquid formulation is put in contact with the target area, tobe substantially retained in the target area, wherein the apparatusdelivers the liquid formulation onto the target area in a volume of fromabout 0.01 ml to about 1 ml.

In several embodiments the a means for delivering photoactive particlesinto small target regions includes apparatus comprising a needle-noseapplicator, a fine tip applicator, a pipette, a dropper, a brush, anapplicator, a module, a capsule, a syringe, and/or a sprayer (e.g., amicro-sprayer such an airbrush). In several embodiments the apparatus iscapable of delivering a volume of from about 1 to about 50 microliters(e.g., 1-10, 10-25, 25-50 microliters) of the formulation on the targetarea such that the surface area of the target area contacted by theformulation is less than about 25 mm². The formulation is liquid in manyembodiments (and includes gelatinous formulations), but also includessolid forms, such as grains, granules, and/or fine powders. In severalembodiments the light source comprises an infrared laser or intensepulsed light (IPL).

In some embodiments, the invention comprises a kit for treating theskin. The kit includes some or all of the following: a formulation ofphotoactive particles (such as nanoparticles and/or chromophores), meansfor delivering the formulation to the skin (e.g., to atrophic regions orother target regions), a light source, and instructions for use. In oneembodiment, an energy source (such as light source) is also included. Insome embodiments a means of removing the formulation of photoactiveparticles from the skin or modifying the distribution of the formulationon the skin is provided. In various embodiments, drugs or othersubstances to be delivered to the dermis and epidermis are providedwhich can either enhance the effects of the treatment, or decrease theside effects caused by partial damage of the epidermis and/or dermis, orboth.

The methods summarized above and set forth in further detail belowdescribe certain actions taken by a practitioner; however, it should beunderstood that they can also include the instruction of those actionsby another party. Thus, actions such as “identifying a target region”can include “instructing the identification of a target region” and“delivering an energy” can include “instructing the delivery of anenergy.”

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is illustrative of schematics depicting certain embodiments ofthe use of formulations for hair removal and acne treatment. Depicted is(A) for hair removal, the plasmonic nanoparticle formulation (black)is 1. applied topically to human skin, 2. delivered deep into thefollicle and washed from the skin surface, 3. irradiated with a clinicallaser at a wavelength resonant to the peak absorption wavelength of theplasmonic particle, and 4. shed from the follicle along with the damagedhair follicle; and (B) for acne treatment, the plasmonic nanoparticleformulation (black) is 1. applied topically to human skin, 2. deliveredspecifically into the sebaceous gland and washed from the skin surface,3. irradiated with a clinical laser at a wavelength resonant to the peakabsorption wavelength of the plasmonic particle, and 4. shed from thetarget site where the accumulated sebum and sebum-producing capabilitiesof the sebaceous gland are destroyed.

FIG. 2 is illustrative of a temperature profile of certain embodimentsof the formulations of plasmonic nanoparticles (SL-001, triangles)provided herein compared to certain embodiments of clinical dyes carbonlotion (circles), meladine spray (diamonds), and indocyanine green(squares), after exposure to 1064 nm, 20 J/cm², 55 ms laser pulses.SL-001 and dyes were equally diluted at 1:1000 from clinicalconcentration (SL-001 1000 O.D., carbon 20-200 mg/ml, meladine 1 mg/ml,ICG 5 mg/ml). n=3, error S.D. of mean.

FIG. 3 is illustrative of hair follicle penetration offluorescently-labeled nanoparticles determined using porcine skinexplants and confocal imaging of certain embodiments of the subjectmatter described herein. Depicted is (A) schematic of treated porcineskin, sectioned and imaged at an angle to the follicle, in two serial 60μm planes: ‘plane 1’ (showing follicle infundibulum) and ‘plane 2’(showing deep follicle); (B) representative confocal images show redfluorescent nanoparticles (548 nm) within superficial and deep follicle,but not in underlying dermis; and (C) red fluorescent nanoparticlesretained in the deep follicle (˜400 μm) at high magnification. Green istissue autofluorescence.

FIG. 4 is illustrative of a hair follicle penetration of plasmonicnanoparticles determined using porcine skin explants and dark fieldimaging. Shown is (A) schematic of treated porcine skin, sectioned andimaged horizontal to the follicle; (B) bright blue plasmonic particlesare visible in a 1.2 mm deep section, and are differentiated from (C)untreated (negative control) porcine skin, where no pigments arevisible.

FIG. 5 depicts clinical observations in live human skin treated withLaser Only (left forearm) or Plasmonic Particles+Laser (right forearm)demonstrates non-specific and specific photothermal damage. (A,B) In thetop panel, human skin was irradiated with 810 nm laser pulses (30 J/cm2,30 ms, 2 passes) alone (A), or after treatment with a formulation of 830nm resonant, Uncoated plasmonic nanoparticles in 20% propylene glycol(B). The plasmonic nanoparticle formulation was applied with 3 minutemassage, and the skin surface wiped with 3 applications of alternativewater and ethanol before laser irradiation. In several embodiments ofthe invention, massage (e.g., hand massage, vibration, mechanicalvibration) can be applied at frequencies of less than 1 kHz, 1 Hz-900Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250 Hz, and any frequenciestherein. At 30 minutes following laser irradiation, non-specificclinical burns were observed in B compared to A, due to significantphotothermal heating of residual, uncoated particles on the skinsurface. (C,D) In the bottom panel, human skin was irradiated with 1064nm laser pulses (40 J/cm2, 55 ms, 3 passes) alone (C), or aftertreatment with a formulation of 1020 nm resonant, Silica-coatedplasmonic nanoparticles in 20% propylene glycol (D). The plasmonicnanoparticle formulation was applied with 3 minute massage, and the skinsurface wiped with 3 applications of alternative water and ethanolbefore laser irradiation. At 30 minutes following laser irradiation, noevidence of burning of the skin or erythema was observed in D or C, asSilica-coated particles could be sufficiently wiped from the skinsurface. Magnified photography of D showed specific photothermal damage(perifollicular erythema and edema) in the nanoparticle-targeted site.

FIG. 6 is a photograph showing nanoparticle-targeted photothermal damagein live human skin treated with a plasmonic nanoparticle formulation andclinical laser. A formulation of 1020 nm resonant, silica-coated (200nm-diameter) plasmonic nanoparticles in 20% propylene glycol and 3minute massage was contacted with live human skin. The procedure wasrepeated 3 times, and skin surface wiped with 3 applications ofalternating water and ethanol to remove residual particles. The treatedskin was irradiated with 1064 nm laser pulses (40 J/cm², 55 ms, 3passes). Following laser irradiation, clinical observation ofperifollicular erythema and edema was visible at hair follicles wherenanoparticles were targeted, but not visible in surrounding ornon-particle-treated tissues.

FIG. 7 is illustrative of a plasmonic nanoparticle formulation deliveryto human skin sebaceous gland. (A) Confocal microscope image of a humanskin biopsy and section, immunostained for Collagen IV basement membrane(blue) and PGP 9.5 nerve marker (green), shows hair follicle (HF) andsebaceous gland (SG) microanatomy. Red is silica nanoparticles (200 nm).(B) Schematic and dark field microscope image of excised human skintreated with plasmonic nanoparticle formulation, then sectioned andimaged horizontal to the follicle. Bright blue plasmonic particles arevisible up to 400 μm deep and within the human sebaceous gland.

FIG. 8 is illustrative of cosmetic formulations of plasmonicnanoparticles for sebaceous gland targeting that include surfactants.Silica-coated nanoparticles (200 nm diameter, 100 O.D.) were formulatedin 20% propylene glycol with the addition of surfactants sodium dodecylsulfate (SDS) or sodium laureth-2 sulfate (SLES), applied to human skinwith massage+ultrasound, and skin was sectioned in horizontal planes fordark field microscopy. (A) Formulations of plasmonic particles in 1%SDS/20% PG penetrated sebaceous gland down to 400 um as in FIG. 7. (B)Formulations of plasmonic particles in 1% SLES/20% PG penetratedsebaceous gland down to 600 um. Inset shows a skin section withoutvisible particles (scale bar 40 um). Sebaceous gland is pseudo-outlined.

FIG. 9 is an image depicting impact of massage vs. ultrasound onnanoparticle targeting to the human follicle and sebaceous gland.Silica-coated nanoparticles (200 nm diameter, 100 O.D.) were formulatedin 1% SDS/20% propylene glycol and applied to human skin with massage orultrasound. Dark field images of horizontal planar sections taken at low(20×) and high (50×) magnification show (A) little to no accumulation ofplasmonic particles into follicle infundibulum after massage alone,compared to (B) follicle infundibulum expansion and significantplasmonic particle accumulation after ultrasound alone. In severalembodiments of the invention, low frequency ultrasound can be applied atfrequencies of 1 kHz to 500 kHz, e.g., 1 kHz-100 kHz, 5 kHz-45 kHz, 20kHz-50 kHz, 30 kHz-40 kHz, 30 kHz, 40 kHz, and any ranges or frequenciestherein.) In several embodiments of the invention, massage (e.g., handmassage, vibration, mechanical vibration) can be applied at frequenciesof less than 1 kHz, 1 Hz-900 Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250Hz, and any frequencies therein.

FIG. 10 depicts an embodiment of the plasmonic nanoparticle cosmeticformulations for sebaceous gland targeting. Plasmonic nanoparticlescomprising different shapes and coatings were formulated in 1% SDS/20%propylene glycol and applied to human skin with massage+ultrasound, andskin was sectioned in horizontal planes for dark field microscopy. (A)Polyethylene glycol (PEG)-coated nanorods (gold, 15×30 nm dimension)were observed within the follicle infundibulum up to 200 um deep (whitearrow). (B) Lower concentration (10 O.D.) Silica-coated nanoplates(silver, 200 nm diameter) were observed up to 600 um deep in thefollicle and in the sebaceous gland (open arrow). Inset shows skinsections without visible particles (scale bar 100 um).

FIG. 11A is illustrative of temperature profiles of certain embodimentsof plasmonic nanoparticle formulations compared to other commercial andresearch chromophores as a function of number of pulses from a 20 J/cm²1064 nm laser (55 ms pulses).

FIG. 11B is illustrative of temperature profiles of certain embodimentsof plasmonic nanoparticle formulations compared to other commercial andresearch chromophores as a function of number of pulses from a 20 J/cm²810 nm laser (30 ms pulses).

FIGS. 12A and 12B are images of embodiments of nanoparticle formulationsin porcine skin.

FIGS. 13A and 13B are images of biopsies taken from in vivo-treatedhuman skin, which were sectioned and immunostained for skin markers,with various embodiments of nanoparticles.

FIG. 14 includes images of an embodiment of a treatment of atrophicscars with a laser and photoactive material delivered to small targetregions of dermal scar tissue.

FIG. 15 is a schematic side view of a composition being distributed froma skin surface to a target in the tissue with a delivery deviceaccording to an embodiment of the invention.

FIG. 16 is a schematic of an experimental process for measuring thedistribution of a composition with various embodiments of deliverydevices in tissue.

FIG. 17 is a table summarizing experimental data measuring theperformance of three embodiments of delivery devices to deliver a singleembodiment of the composition to various depths in tissue.

FIG. 18 illustrates images of the delivery of a composition in skinmodels with various embodiments of delivery devices.

FIG. 19 is illustrative of a graph of experimental measurements ofpercentage of composition delivery to an area at a depth in tissue withvarious delivery devices according to embodiments of the invention.

FIG. 20 is illustrative of a graph of experimental measurements ofpercentage of composition delivery to an area at a depth in tissue withvarious delivery devices according to embodiments of the invention.

FIG. 21 is illustrative of images taken of tissue sections atapproximately 720 microns deep of composition delivery within tissuesamples using various embodiments of delivery devices. For the deliverydevice embodiment of a vibration device (Vibraderm) plus acoustichorn/sonotrode ultrasound energy, there is a markedly increase in thetotal amount of composition delivered (100) and number of follicles thatthe composition is delivered to over the other delivery embodimentsalone. At levels of approximately 720 microns, there is also asignificant increase in delivery of the composition with vibrationdevice (Vibraderm) plus 30-40 kHz ultrasound (flat transducer) over thevibration device alone.

FIG. 22 is illustrative of images taken of tissue sections atapproximately 1060 microns deep of composition delivery within tissuesamples using various embodiments of delivery devices. For the deliverydevice embodiment of a vibration device plus acoustic horn/sonotrodeultrasound energy, there is a markedly increase in the total amount ofcomposition delivered (100) and number of follicles that the compositionis delivered to over the other delivery embodiments alone. At this depthof approximately 1060 microns, the amount delivery of the composition bythe other delivery device embodiments has significantly decreased.

DETAILED DESCRIPTION

The biology of physiological and pathophysiological tissue growth andremodeling, and alterations in cell morphology is more complex thangenerally appreciated, involving an interacting network of biologicalcompounds, physical forces, and cell types.

An object of the subject matter described herein is to providecompositions, methods and systems for noninvasive and minimally-invasivetreatment of skin and underlying tissues, or other accessible tissuespaces with the use of photoactive compounds (including but not limitedto photoactive particles such as nanoparticle, plasmonic nanoparticles,etc.). In some embodiments, the invention describes the development andutilization of compositions containing photoactive materials (e.g.,nanoparticles and other materials) for the treatment of small targetregions of skin including acne scars and other skin conditions. In oneembodiment, such compositions are generally applied topically, throughan apparatus that provides the composition in a form suitable forcontact with and retention at a target region of skin in a manner thatencompasses irradiating the skin with light (e.g., electromagneticradiation) having a wavelength sufficient to ablate or otherwise damagethe target region of skin and cause remodeling of the skin tissue. Invarious embodiments, the treatment includes, but is not limited to, hairremoval, hair growth and regrowth, and skin rejuvenation or resurfacing,acne removal or reduction, wrinkle reduction, pore reduction, ablationof cellulite and other dermal lipid depositions, wart and fungusremoval, thinning or removal of scars including hypertrophic scars andkeloids, abnormal pigmentation (such as port wine stains), tattooremoval, and skin inconsistencies (e.g. in texture, color, tone,elasticity, hydration, and including sun spots, age spots, freckles, andother inconsistencies). Other therapeutic or preventative methodsinclude but are not limited to treatment of hyperhidrosis, anhidrosis,Frey's Syndrome (gustatory sweating), Homer's Syndrome, and RossSyndrome, actinic keratosis, sebhorreic keratosis, keratosisfollicularis, dermatitis, vitiligo, pityriasis, psoriasis, lichenplanus, eczema, alopecia, psoriasis, malignant or non-malignant skintumors, onychomycosis, sebhorreic dermatitis, atopic dermatitis, contactdermatitis, herpes simplex, Human papillomavirus (HPV), anddermatophytosis.

Unless explained otherwise, all technical and scientific terms usedherein have the same meaning as commonly understood to one of ordinaryskill in the art to which this disclosure belongs. Although methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present disclosure, suitable methods andmaterials are described herein. The materials, methods, and examples areillustrative only and not intended to be limiting. Other features of thedisclosure are apparent from the following detailed description and theclaims.

“Administer” and “administration” as used herein, include providing orcausing the provision of a material to a subject, such as by a topical,subdermal, subcutaneous, intradermal, enteral, parenteral, rectal,nasal, intravenous, intramuscularly, intraperitoneal, or other route.

A “carrier suitable for administration” to a subject is any materialthat is physiologically compatible with a topical or route ofadministration to a desired vertebrate subject. Carriers can includesolid-based, dry materials for formulation; or the carrier can includeliquid or gel-based materials for formulations into liquid or gel forms.The specific type of carrier, as well as the final formulation depends,in part, upon the selected route(s) of administration and the type ofproduct.

A “comparable amount” is an amount that is measurably similar to a givenreference or standard.

The “components” of a formulation include any products or compoundsassociated with or contained within it.

An “effective dose”, “effective amount” or “therapeutic amount” is anamount sufficient to elicit the desired pharmacological, cosmetic ortherapeutic effects, thus resulting in effective prevention or treatmentof a disease or disorder, or providing a benefit in a vertebratesubject.

A “therapeutic effect” or “therapeutically desirable effect” refers to achange in a domain or region being treated such that it exhibits signsof being effected in the manner desired, e.g., cancer treatment causesthe destruction of tumor cells or halts the growth of tumor cells, acnetreatment causes a decrease in the number and/or severity of blemishes,hair removal treatment leads to evident hair loss, or wrinkle reductiontreatment causes wrinkles to disappear.

An “isolated” biological component (such as a nucleic acid molecule,protein, or cell) has been substantially separated or purified away fromother biological components in which the component was produced,including any other proteins, lipids, carbohydrates, and othercomponents.

A “nanoparticle”, as used herein, refers generally to a particle havingat least one of its dimensions from about 0.1 nm to about 9000 nm.(e.g., 1 nm-500 nm, 10 nm-300 nm, 100 nm-500 nm.).

A “subject” or “patient” as used herein is any vertebrate species.

As used herein, a “substantially pure” or “substantially isolated”compound is substantially free of one or more other compounds.

A “target tissue” includes a region of an organism to which a physicalor chemical force or change is desired. As described herein, variousembodiments of target tissues for acne treatment include a sebaceousgland, while various embodiments of target tissues for hair removalinclude a pilosebaceous unit, a hair infundibulum, a hair follicle, or anon-follicular epidermis. Target tissues for sweat or hyperhidrosisinclude a sweat gland. A “region” of a target tissue includes one ormore components of the tissue. In some embodiments, target tissueregions include the stem cell niche, bulge, sebaceous gland, dermalpapilla, cortex, cuticle, inner root sheath, outer root sheath, medulla,Huxley layer, Henle layer or arrector pili muscle. A “domain” of atarget tissue region includes basement membrane, extracellular matrix,cell-surface proteins, unbound proteins/analytes, glycomatrices,glycoproteins, or lipid bilayer.

A compound that is “substantially free” of some additional contents islargely or wholly without said contents.

A “plasmonic nanoparticle” is a nanometer-sized metallic structurewithin which localized surface plasmons are excited by light. Thesesurface plasmons are surface electromagnetic waves that propagate in adirection parallel to the metal/dielectric interface (e.g., metal/air ormetal/water).

A “light-absorbing nanomaterial” includes a nanomaterial capable ofdemonstrating a quantum size effect.

As described herein, provided are compositions that contain plasmonicnanoparticles to induce selective thermomodulation in a target tissue.

Plasmonic Nanoparticles.

In various embodiments, a composition comprises plasmonic nanoparticles.In various embodiments, such compositions contain from about 2 to about1×10¹⁸ or up to 10²³ nanoparticles (e.g., 10⁹ to about 10¹⁶nanoparticles), such as 10², 10³, 10⁴, 10⁵, 10⁶, 10⁷, 10⁸, 10⁹, 10¹⁰,10¹¹, 10¹², 10¹³, 10¹⁴, 10¹⁵, 10¹⁶, 10¹⁷, or 10¹⁸ particles, includingsuch as ranges between about 10⁹ to 10²³, 10⁹ to 10¹⁸, 10⁹ to 10¹⁶, 10⁹to 10¹⁵, 10⁹ to 10¹⁴, 10⁹ to 10¹³, 10⁹ to 10¹², 10⁹ to 10¹¹, 10 10⁹ to10¹¹ particles per ml. In one embodiment, the range is between about 10⁵to 10¹⁸ per ml of composition or solution. In one embodiment, the rangeis between about 10⁹ to 10¹⁶ per ml of composition or solution Thenumbers of particles may include the number, or any range of any numbersdisclosed. In some embodiments, a concentration of particles isexpressed as the number of particles per milliliter of, for example, thesolution. In one embodiment, the compositions contain about 10¹¹ to 10¹³particles so that the amount of particles localized to an effective 1 mltreatment volumes is from 10⁹ to 10¹¹. In various embodiments, thecompositions contain nanoparticles in a concentration of from about 1O.D. to about 10,000 O.D. For embodiments wherein a greaterconcentration of nanoparticles to a target region is desired,compositions contain particle concentrations with optical densities of,for example, 10 O.D.-5000 O.D. more specifically 100 O.D.-1000 O.D., oroptical densities greater than 1,000 O.D. In certain embodiments whereinincreased concentration of nanoparticles to a target region is desired,compositions contain particle concentrations with optical densities(O.D.) of 10 O.D.-1000 O.D., or optical densities greater than 1,000O.D. In some embodiments, the optical density of a composition is any of100 O.D., plus or minus 10%, plus or minus 5%, and/or plus or minus 1%.In some embodiments, the optical density of a composition is 100 O.D. to10,000 O.D. e.g., 1000 O.D., plus or minus 10%, plus or minus 5%, and/orplus or minus 1%. In some embodiments these correspond to concentrationsof about 1-10% w/w (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10%, orranges within these numbers) or more of nanoparticles. Determination ofO.D. units in a composition is determined using devices and analysesknown in the art.

Nanoparticles may be homogenous or heterogeneous in size and othercharacteristics. The size of the nanoparticle is generally about 0.1 nmto about 50,000 nm (e.g., about 0.1 nm to about 5,000 nm) in at leastone dimension. Some variation in the size of a population ofnanoparticles is to be expected. For example, the variation might beless than 0.01%, 0.1%, 0.5%, 1%, 5%, 10%, 15%, 25%, 50%, 75%, 100%, 200%or greater than 200%, or any range between any numbers therein (e.g.,0.01%-200%, 0.1%-10%, 15%-75%, 0.1%-100%, etc.). In certain embodimentswhere optimal plasmonic resonance is desired, a particle size in therange of from about 10 nm to about 100 nm is provided. Alternatively, inembodiments where enhanced penetration of the nanoparticles into atarget tissue region such as a hair follicle is desired, a particle sizein the range of from about 100 nm to about 1000 nm is provided.Modulation of particle size present in the composition is also a usefulmeans of concentrating the composition in a target domain. Further, asdescribed herein, nanoparticles having a size range of from about 10 nmto about 100 nm can be used as component of a larger molecularstructure, generally in the range of from about 100 nm to about 1000 nm.For example, the plasmonic nanoparticle can be surface coated toincrease its size, embedded into an acceptable carrier, or it can becross-linked or aggregated to other particles, or to other materials,that generate a larger particle. In certain embodiments where at leastone dimension of at least one nanoparticle within a solution ofplasmonic nanoparticles is below 50-100 nm, the nanoparticle surface canbe coated with a matrix (e.g. silica) of 10-100 nm thickness or more inorder to increase that dimension or particle to 50-100 nm or more. Thisincreased dimension size can increase the delivery of all nanoparticlesto a target region (e.g., hair follicle) and limit delivery tonon-target region (e.g. dermis). In one embodiment, the inventioncomprises a composition comprising at least about 1 O.D. (e.g., at least10 O.D.) of coated plasmonic nanoparticles (e.g., comprising silica orpolyethylene glycol (PEG)) having a mean length in at least onedimension greater than about 30 nanometers, wherein the coatednanoparticles are formulated in an acceptable carrier to be effective ininduction of selective thermoablation in a target tissue region withwhich the composition is contacted, wherein the affinity of the coatednanoparticles for the target tissue region is substantially greater thanthe affinity of the coated nanoparticles for a non-target tissue region.

Some considerations when generating embodiments of nanoparticlesinclude: 1) the zeta potential (positive, negative, or neutral) andcharge density of the particles and resulting compositions; 2) thehydrophilicity/hydrophobicity of the particles and resultingcompositions; 3) the presence of an adsorption layer (e.g., a particleslippage plane); and 4) target cell adhesion properties. Nanoparticlesurfaces can be functionalized with thiolated moieties having negative,positive, or neutral charges (e.g. carboxylic acid, amine, hydroxyls) atvarious ratios. Moreover, anion-mediated surface coating (e.g. acrylate,citrate, and others), surfactant coating (e.g., sodium dodecyl sulfate,sodium laureth 2-sulfate, ammonium lauryl sulfate, sodiumoctech-1/deceth-1 sulfate, lecithin and other surfactants includingcetyl trimethylammonium bromide (CTAB), lipids, peptides), orprotein/peptide coatings (e.g. albumin, ovalbumin, egg protein, milkprotein, other food, plant, animal, bacteria, yeast, orrecombinantly-derived protein) can be employed. Block-copolymers arealso useful. Further, one will appreciate the utility of any othercompound or material that adheres to the surface of light-absorbingparticles to promote or deter specific molecular interactions andimprove particle entry into pores or follicles. In some embodiments, theparticle surface is unmodified. Modulation of hydrophilicity versushydrophobicity is performed by modifying nanoparticle surfaces withchemistries known in the art, including thiols, dithiolane, silanes,isothiocyanates, short polymers (e.g., PEG), or functionalizedhydrocarbons. Polymer chains (e.g., biopolymers such as proteins,polysaccharides, lipids, and hybrids thereof; synthetic polymers such aspolyethyleneglycol, PLGA, and others; and biopolymer-synthetic hybrids)of different lengths and packing density are useful to vary theadsorption layer/slippage plane of particles.

Optical absorption. In various embodiments, nanoparticles have opticalabsorption qualities of about 10 nm to about 10,000 nm, e.g., 100-500nm, 500-750 nm, 600-900 nm, 700-1,000 nm, 800-1,200 nm, or 500-2,000 nm.In specific embodiments, the nanoparticles have optical absorptionuseful to excitation by standard laser devices or other light sources.For example, nanoparticles absorb at wavelengths of about 755 nm(alexandrite lasers), in the range of about 800-810 nm (diode lasers),or about 1064 nm (Nd: YAG lasers). Similarly, the nanoparticles absorbintense pulsed light (IPL), e.g., at a range of about 500 nm to about1200 nm.

Assembly. In various embodiments, the nanoparticles can contain acollection of unassembled nanoparticles. By “unassembled” nanoparticlesit is meant that nanoparticles in such a collection are not bound toeach other through a physical force or chemical bond either directly(particle-particle) or indirectly through some intermediary (e.g.particle-cell-particle, particle-protein-particle,particle-analyte-particle). In other embodiments, the nanoparticlecompositions are assembled into ordered arrays. In particular, suchordered arrays can include any three dimensional array. In someembodiments, only a portion of the nanoparticles are assembled, e.g., 5,10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 86, 90, 95,99% or greater than 99% of the nanoparticles are assembled in an orderedarray. The nanoparticles are assembled by a van der Walls attraction, aLondon force, a hydrogen bond, a dipole-dipole interaction, or acovalent bond, or a combination thereof.

“Ordered array”-“Ordered arrays” can take the form of a macrostructurefrom individual parts that may be patterned or unpatterned in the formof spheres, colloids, beads, ovals, squares, rectangles, fibers, wires,rods, shells, thin films, or planar surface. In contrast, a “disorderedarray” lacks substantial macrostructure.

Geometrically tuned nanostructures. In various embodiments, theparticles are formable in all shapes currently known or to be createdthat absorb light and generate a plasmon resonance at a peak-wavelengthor composition of wavelengths from 200 nm to 10,000 nm. In non-limitingexamples, the nanoparticles are shaped as spheres, ovals, cylinders,squares, rectangles, rods, stars, tubes, pyramids, stars, prisms,triangles, branches, plates or comprised of a planar surface. Innon-limiting examples, the plasmonic particles comprise nanoplates,solid nanoshells, hollow nanoshells nanorods, nanorice, nanospheres,nanofibers, nanowires, nanopyramids, nanoprisms, or a combinationthereof. Plasmonic particles present in the composition comprise asubstantial amount of geometrically-tuned nanostructures defined as 5,10, 15, 25, 50, 75, 80, 85, 90, 95, 98, 99, 99.9 or greater than 99.9%of particles.

Composition. In various embodiments, the nanoparticle is a metal (e.g.,gold, silver), metallic composite (e.g., silver and silica, gold andsilica), metal oxide (e.g. iron oxide, titanium oxide), metallic salt(e.g., potassium oxalate, strontium chloride), intermetallic (e.g.,titanium aluminide, alnico), electric conductor (e.g., copper,aluminum), electric superconductor (e.g., yttrium barium copper oxide,bismuth strontium calcium copper oxide), electric semiconductor (e.g.,silicon, germanium), dielectric (e.g., silica, plastic), or quantum dot(e.g., zinc sulfide, cadmium selenium). In non-limiting examples, thematerials are gold, silver, nickel, platinum, titanium, palladium,silicon, galadium. Alternatively, the nanoparticle contains a compositeincluding multiple metals (e.g., alloy), a metal and a dielectric, ametal and a semiconductor, or a metal, semiconductor and dielectric.

Coating. In some embodiments, the composition contains coated particles.

Type of Material Properties Examples of Materials Bio- Moiety withaffinity or avidity Antibody, peptide, phage, recognitive for asubstrate or analyte DNA, RNA material Bioactive Moiety (e.g., protein,analyte) Growth factor (e.g. VEGF), material that interrogates ormodulates cytokine, cell surface the activity of biologic entityreceptors, receptor ligands, or cell G-protein, kinase/ phosphataseBiological Material that is sourced from albumin, ovalbumin, eggmaterial living matter protein, milk protein, other food, plant, animal,bacteria, yeast, or recombinantly- derived protein; peptides; enzymes,lipids, fatty acids, sugars Biocide Material that is active in Syntheticor natural material killing, destroying, or pesticides, synthetic ordisturbing biological natural anti-microbials matter Dielectric Aninsulator that may be Silicon, doped materials polarized by an electricfield semiconductors Chemo- Material that is able to interact Receptor,receptor ligand, recognitive with a moiety for binding, chemicalmolecule material biological or chemical reactions Chemical Materialthat causes the Aldehyde, halogens, metals active transformation of asubstance material Polymer/ Long chain molecule (linear PLGA, PEG, PEO,dendrimer or branched, block or co- polystyrene, carboxylate block)styrene, rubbers, nylons, silicones, polysaccharides Environ- Surfacemolecule that changes Ph sensitive bond, light mentally by itsenvironment (e.g. acid) sensitive bond, heat sensitive sensitive bond,enzyme sensitive polymer bond, hydrolytic bond Hydrogel Polymer withhigh Synthetic 2-hydroxyethyl hydrophilicity and water metacrylate(HEMA)-based, “ordering” capacity polyethylene glycol (PEG)- based,PLGA, PEG- diacrylate; Natural ionic gels, alginate, gelatin, hyaluronicacids, fibrin Metal Thin metal coating to achieve Gold, silver, nickel,copper, improved resonance and/or platinum, titanium, functionalizationcapacity chromium, palladium. Semi- Semiconductor layer or core Siliconand galadium. conductors that enhance Plasmon resonance PolymerFluorophore cross linked to a Fluorescein, rhodamine, containing apolymer coat or directly to the Cy5, Cy5.5, Cy7, Alexa fluorescentsurface of the particle dyes, Bodipy dyes marker Matrix Matrix coatingthat increases Silica, polyvinyl pyrrolidone, solubility ofnanoparticles polysulfone, polyacrylamide, and/or reduces “stickiness”polyethylene glycol, to biological structures polystyrene cellulose,pplyquaterniums, lipids, surfactants, carbopol.

Biological molecules. In various embodiments, the composition maycontain a peptide, a nucleic acid, a protein, or an antibody. Forexample a protein, antibody, peptide, or nucleic acid that binds aprotein of a follicular stem cell (e.g., keratin 15), a protein,glycomatrix, or lipid on the surface of a cell or stem cell, a protein,peptide, glycomatrix of the extracellular matrix or basement membrane.

Charged moieties. In various embodiments, the coated nanoparticles maycontain charged moieties whereby those charges mediate enhanced ordiminished binding to components within or outside the hair follicle viaelectrostatic or chemical interactions.

Class of Moiety Properties Examples of Moieties Polar moieties Neutralcharge but increases Hydroxyl groups, hydrophilicity in waterisothiocyanates Non-polar moieties Increases hydrophobicity and orHydrocarbons, myristoylated improves solubility compounds, silanes,isothiocyanates Charged moieties Functional surface modificationsAmines, carboxylic acids, that change the zeta potential, hydroxylsisoelectric point, or pKa, and impact adsorption/binding tocomplementary charge compounds Ionic moieties Surface groups that have asingle Ammonium salts, chloride ion salts Basic moieties Groups thatdonate a hydrogen Amides, hydroxides, metal ions oxides, fluoride Acidicmoieties Moieties that accept hydrogen Carboxylic acids, sulfonic ionsacids, mineral acids Oxidative moieties Moieties that oxidize Manganeseions, reactive oxygen species Hydrophobic moieties Moieties that improvesolubility in Hydrocarbons, myristoylated non-aqueous solution and/orcompounds, silanes improve adsorption on the skin within a hair follicleHydrophilic moieties Moieties that are water-loving and PEG, PEO, PLGAprevent adsorption Agnostic moieties Moieties that bind a target cell,Antibodies, peptides, proteins structure, or protein of interestAntagonistic moieties Moieties that block the binding to Antibodies,peptides, proteins a target of interest Reactive moieties Moieties thatreact with biological Aldehydes or non-biological components with aresulting change in structure on the targetDescription of Target Tissues.

Topical and Dermatological Applications. In some embodiments, targettissues for topical and dermatological applications include the surfaceof the skin, the epidermis and the dermis. Diseases or conditionssuitable for treatment with topical and dermatological applicationsinclude acne, warts, fungal infections, psoriasis, scar removal, hairremoval, hair growth, reduction of hypertrophic scars or keloids, skininconsistencies (e.g. texture, color, tone, elasticity, hydration), andmalignant or non-malignant skin tumors.

As used herein, the term “acne” includes acne vulgaris as well as otherforms of acne and related cutaneous conditions, including acneaestivalis, acne conglobata, acne cosmetic, acne fulminans, acnekeloidalisnuchae, acne mechanica, acne miliarisnecrotica, acnenecrotica, chloracne, drug-induced acne, excoriated acne, halogen acne,lupus miliaris disseminates faciei, pomade acne, tar acne, and tropicalacne.

Subdermal Applications. In some embodiments, target tissues forsubdermal applications include the adipose tissue and connective tissuebelow the integumentary system. Diseases or conditions suitable fortreatment with subdermatological applications include wrinkles andtattoos. Other applications with photoactive particles (e.g., plasmonicnanoparticles) include skin rejuvenation and/or resurfacing, the removalor reduction of stretch marks and fat ablation.

In some embodiments, a specific region of the target tissue that can betreated with the photoactive particles (e.g., plasmonic nanoparticles)descried herein is a hair follicle, a sebaceous gland, a merocrine sweatgland, an apocrine sweat gland, or an arrector pili muscle, within whicha specific domain is targeted. For example, the bulge region of the hairfollicle is targeted. Because in one embodiment the nanoparticles areuseful to thermally ablate hair follicle stem cells for hair removal,regions containing hair follicle stem cells are of particular interestfor targeting. Thus, the target tissue region may include a stem cellniche, bulge, sebaceous gland, dermal papilla, cortex, cuticle, innerroot sheath, outer root sheath, medulla, Huxley layer, Henle layer orarrector pili muscle. Each of these regions may contain cells, stemcells, basement membrane, extracellular matrix, growth factors,analytes, or other biologic components that mediate hair folliclerejuvenation. Disruption or destruction of these components would have atherapeutic effect, e.g. slow or stop the processes that mediate hairregrowth, prevent the secretion of sebum from the sebaceous gland,damage or deter tumor cells, reduce the appearance of wrinkles.Structures can also be targeted that are in close proximity to a desiredtarget for ablation, especially when capable of conducting heateffectively.

Localization Domains. In one embodiment, compositions containingnanoparticles (e.g., plasmonic or non-plasmonic nanoparticles) thatpreferentially localize to a domain of a target tissue region of amammalian subject to whom the composition is administered.

Targeting moieties. In some embodiments, nanoparticles (e.g., plasmonicor non-plasmonic nanoparticles) can be engineered to selectively bind toa domain of the target tissue. For example, the nanoparticles areoperably linked to the domain via a biologic moiety, in order toeffectively target the nanoparticles to the target tissue domain.Preferably, the moiety contains a component of a stem cell, a progenitorcell, an extracellular matrix component, a basement membrane component,a hair shaft component, a follicular epithelial component, or anon-follicular epidermal component. Biological moieties include proteinssuch as cell surface receptors, glycoproteins or extracellular matrixproteins, as well as carbohydrates, analytes, or nucleic acids (DNA,RNA) as well as membrane components (lipid bilayer components,microsomes).

Delocalization Domains. In some embodiments, nanoparticles (e.g.,plasmonic or non-plasmonic nanoparticles) present in the compositionpreferentially delocalize away from a domain of a target tissue region.Delocalization domains include specific regions of a tissue into whichnanoparticles do not substantially aggregate, or alternatively, areremoved from the domain more effectively. The delocalization domain,according to several embodiments, is a non-follicular epidermis, dermis,a component of a hair follicle (e.g., a hair stem cell, a stem cellniche, a bulge, a sebaceous gland, a dermal papilla, a cortex, acuticle, an inner root sheath, an outer root sheath, a medulla, a Huxleylayer, a Henle layer, an arrector pili muscle), a hair follicleinfundibulum, a sebaceous gland, a component of a sebaceous gland, asebocyte, a component of a sebocyte, or sebum

Energy sources. Provided herein are various embodiments of energysources to, for example, apply to or otherwise activate the photoactiveparticles. These include, but are not limited to, surface plasmonresonance excitation sources (e.g., nonlinear excitation surface plasmonresonance sources), various light sources and optical sources. Variousembodiments of light sources include a laser (ion laser, semiconductorlaser, Q-switched laser, free-running laser, or fiber laser), lightemitting diode, lamp, the sun, a fluorescent light source or anelectroluminescent light source. In several embodiments, the energysource is capable of emitting radiation at a wavelength from about 100,200, 300, 400, 500, 1000, 2000, 5000 nm to about 10,000 nm or more. Thesurface plasmon resonance excitation sources (e.g., nonlinear excitationsurface plasmon resonance source) is capable of emitting electromagneticradiation, ultrasound, thermal energy, electrical energy, magneticenergy, or electrostatic energy. For example, the energy is radiation atan intensity from about 0.00005 mW/cm² to about 1000 TW/cm². The optimumintensity is chosen to induce high thermal gradients from plasmonicnanoparticles in regions from about 10 microns to hundreds of microns inthe surrounding tissue, but has minimal residual effect on heatingtissue in which particles do not reside within a radius of about 100microns or more from the nanoparticle. In certain embodiments, adifferential heat gradient between the target tissue region and othertissue regions (e.g., the skin) is greater than 2-fold, 3-fold, 5-fold,10-fold, 15-fold, 20-fold, 50-fold, 100-fold, or greater than 100 fold.With respect to nonlinear embodiments, at resonance wavelengthsplasmonic nanoparticles can act as antennas, providing a nonlinearexcitation at peak resonance or, in other words, an enhanced extinctioncross section for a given physical cross-section of material whencompared to non-plasmonic photoactive materials of the same dimension.Thus, in several embodiments, plasmonic materials may be able to pullmore energy from delocalized electromagnetic waves surrounding thematerial at peak resonance than non-plasmonic photoactive material ofthe same dimension.

The energy can be tuned by monitoring thermal heat gradients on thesurface of the skin with a thermal/infrared camera. As demonstratedherein, the methods and systems of the present disclosure providesuperior efficacy when a surface plasmon is generated on thenanoparticles by the action of the radiation. Typically, the plasmon isgenerated in a one-photon mode or, alternatively, a two-photon mode, amulti-photon mode, a step-wise mode, or an up-conversion mode.

Delivery of radiation. In some embodiments, physical means of deliveryof the light energy (e.g., laser, flash lamp, intense pulse, etc.) tothe surface plasmon resonance excitation (e.g., nonlinear excitationsurface plasmon resonance source) to the target tissue region include afiber, waveguide, a contact tip, a microlens array, a digitalmicromirror array (DMA), or a combination thereof.

In some embodiments, optical sources include a CW optical source or apulsed optical source, which may be a single wavelength polarized (or,alternatively, unpolarized) optical source capable of emitting radiationat a frequency from about 200 nm to about 10,000 nm. Alternatively, theoptical source is a multiple wavelength polarized (or, alternatively,unpolarized) optical source capable of emitting radiation at awavelength from about 200 nm to about 10,000 nm. The pulsed opticalsource is generally capable of emitting pulsed radiation at a frequencyfrom about 1 Hz to about 1 THz. The pulsed optical source is capable ofa pulse less than a millisecond, microsecond, nanosecond, picoseconds,or femtosecond in duration. For example, a source emitting radiation ata wavelength of 755 nm is operated in pulse mode such that the emittedradiation is pulsed at a duration of 0.25-300 milliseconds (ms) perpulse, with a pulse frequency of 1-10 Hz. In another example, radiationemitted at a wavelength of 810 nm is pulsed at 5-100 ms with a frequencyof 1-10 Hz. In a further example, a source emitting radiation at awavelength of 1064 nm is pulsed at 0.25-300 ms at a frequency of 1-10Hz. In yet another example, a source emitting intense pulsed light at awavelength of 530-1200 nm is pulsed at 0.5-300 ms at a frequency of 1-10Hz. The optical source may be coupled to a skin surface cooling deviceto reduce heating of particles or structures on the skin surface andfocus heating to components within follicles or tissue structures atdeeper layers. In some embodiments, pulse widths range from 0.1 ms to 1m, 1 ms-10 ms, 10 ms-100 ms, 100 ms-1000 ms, greater than 1000 ms.

Nanoparticle-containing compositions. In order to provide optimal dermalpenetration into the target tissue, photoactive particles (e.g.,plasmonic nanoparticles) in certain embodiments are formulated invarious compositions. In one embodiment, the nanoparticles areformulated in compositions containing 1-10% v/v surfactants (e.g. sodiumdodecyl sulfate, sodium laureth 2-sulfate, ammonium lauryl sulfate,sodium octech-1/deceth-1 sulfate). Surfactants disrupt and emulsifysebum or other hydrophobic fluids to enable improved targeting ofhydrophilic nanoparticles to the hair follicle, infundibulum, sebaceousgland, or other regions of the skin. Surfactants also lower the freeenergy necessary to deliver hydrophilic nanoparticles into smallhydrophobic crevices such as the space between the hair shaft andfollicle or into the sebaceous gland. Nanoparticle-containingcompositions may also include emulsions at various concentrations (1-20%w/v) in aqueous solutions, silicone/oil solvents, polypropylene gel,propylene glycol or creams (e.g. comprising alcohols, oils, paraffins,colloidal silicas). In other embodiments, the formulation contains adegradable or non-degradable polymer, e.g., syntheticpolylactide/co-glycolide co-polymer, porous lauryllactame/caprolactamenylon co-polymer, hydroxyethylcellulose, polyelectrolyte monolayers, oralternatively, in natural hydrogels such as hyaluronic acid, gelatin andothers. In further embodiments, a hydrogel PLGA, PEG-acrylate isincluded in the formulation. Alternatively, a matrix component such assilica, polystyrene or polyethylene glycol is provided in theformulation. Other formulations include components of surfactants, alipid bilayer, a liposome, microsome, polymersomes, or a polymermicrocapsules. A nanoparticle may comprise a larger micron-sizedparticle.

Effective doses. In various embodiments, an effective dose of thenanoparticle-containing compositions includes an amount of particlesrequired, in some aspects, to generate an effective heat gradient in atarget tissue region, such that a portion of the target tissue region isacted upon by thermal energy from excited nanoparticles. A “minimaleffective dose” is the smallest number or lowest concentration ofnanoparticles in a composition that are effective to achieve the desiredbiological, physical and/or therapeutic effect(s). In some embodiments,the photoactive particles (e.g., plasmonic nanoparticles) have anoptical density of 10 O.D.-1,000 O.D. (e.g., 10-100 O.D, 50-200 O.D,20-300 O.D.) at one or a plurality of peak resonance wavelengths.

Cosmetically acceptable carriers. In some embodiments, provided arecosmetic or pharmaceutical compositions with a plurality of photoactiveparticles (e.g., plasmonic nanoparticles) and a cosmetically orpharmaceutically acceptable carrier. Generally, the carrier andcomposition must be suitable for topical administration to the skin of amammalian subject, such that the photoactive particles (e.g., plasmonicnanoparticles) are present in an effective amount for selectivethermomodulation of a component of the skin. In one embodiment, thenanoparticles are formulated with a carrier containing 1-10% v/vsurfactants (e.g. sodium dodecyl sulfate, sodium laureth 2-sulfate,ammonium lauryl sulfate, sodium octech-1/deceth-1 sulfate) to enabledisruption of the epidermal skin barrier, emulsify sebum, improve mixingof hydrophilic nanoparticles with hydrophobic solutions, and reduceentropic barriers to delivering hydrophilic particles to hydrophobicregions of the skin (e.g. between the hair shaft and surrounding sheathor follicle). In some embodiments, the carrier contains a polar ornon-polar solvent. For example, suitable solvents include alcohols(e.g., n-Butanol, isopropanol, n-Propanol, Ethanol, Methanol),hydrocarbons (e.g., pentane, cyclopentane, hexane, cyclohexane, benzene,toluene, 1,4-Dioxane), chloroform, Diethyl-ether, water, water withpropylene glycol, acids (e.g., acetic acid, formic acid), bases,acetone, isooctanes, dimethyl sulfoxide (DMSO), dimethylformamide (DMF),acetonitrile (MeCN), tetrahydrofuran (THF), dichloromethane (DCM),ethylacetate, tetramethylammonium hydroxide, isopropanol, and others. Inother embodiments, a stabilizing agent such as antioxidants, preventingunwanted oxidation of materials, sequestrants, forming chelate complexesand inactivating traces of metal ions that would otherwise act ascatalysts, emulsifiers, ionic or non-ionic surfactants, cholesterol orphospholipids, for stabilization of emulsions (e.g. egg yolk lecithin,sodium stearoyllactylate, sodium bis(2-ethylhexyl-sulfosuccinate (AOT)),ultraviolet stabilizers, protecting materials, especially plastics, fromharmful effects of ultraviolet radiation is provided. In furtherembodiments, a composition with a cosmetically acceptable carrier isgenerated such that the nanoparticles are substantially in a suspension.

Other components are also optionally included, including an emulsion,polymer, hydrogel, matrix, lipid bilayer, liposome, microsome,polymersome, or polymer microcapsule. Additionally, inclusion of adetectable colorant (e.g., a pigment), a fragrance, a moisturizer,and/or a skin protectant is optional. In some examples, the formulationhas a viscosity of above, below or within 0.1-10,000 (e.g., 5e⁻⁴×10³,1,000), as measured in millipascal-seconds (mPa·s).

In some embodiments, nanoparticle quantities per milliliter in acomposition are subject to modification for specific binding and canrange from 10⁹ to 10¹⁸ particles but generally about 10¹¹ to 10¹³nanoparticles per milliliter. Nanoparticle quantities per milliliter ina formulation are subject to modification for specific binding butgenerally up to about 10²³ nanoparticles per milliliter. In certainembodiments wherein increased concentration of nanoparticles to a targetregion is desired, compositions contain particle concentrations withoptical densities of 10 O.D.-1000 O.D. (e.g., 10-100 O.D, 50-200 O.D,20-300 O.D.), or optical densities greater than 1,000 O.D. (e.g.,1,200-1,500, 2,000) In some embodiments these correspond toconcentrations of about 0.1-10% w/w or more of nanoparticles.

In one embodiment, prior to application of nanoparticle formulations(e.g., photoactive nanoparticles, such as plasmonic nanoparticles), skinand hair follicles can be pre-treated to increase the delivery ofnanoparticles to a target region. In some embodiments, hair shafts arecut or removed via shaving, waxing, sugaring, cyanoacrylate surfacepeels, calcium thioglycolate treatment, or other techniques to removethe hair shaft and/or hair follicle plugs and create a void whereinnanoparticles can accumulate. Orifices of active or inactive folliclescan be blocked by plugs formed of corneocytes and/or other material(e.g. cell debris, soot, hydrocarbons, cosmetics). In some embodimentspre-treatment with surface exfoliation including mechanical exfoliation(e.g., salt glow or microdermabrasion) and chemical exfoliation (e.g.,enzymes, alphahydroxy acids, or betahydroxy acids) removes plugs fromthe orifice of follicles to increase the targeting of nanoparticleformulations to target regions within the hair follicle.

In some embodiments, the nanoparticle formulations (e.g., photoactivenanoparticles, such as plasmonic nanoparticles) are formulated forapplication by a sponge applicator, cloth applicator, direct contact viaa hand or gloved hand, spray, aerosol, vacuum suction, high pressure airflow, or high pressure liquid flow, roller, brush, planar surface,semi-planar surface, wax, ultrasound and other sonic forces (e.g., lowfrequency ultrasound via a transducer, acoustic horn, or sonotrode),mechanical vibrations, physical manipulation, hair shaft manipulation(including pulling, massaging), physical force, electrophoresis,iontophoresis, thermal manipulation, and other treatments. In severalembodiments of the invention, massage (e.g., hand massage, vibration,mechanical vibration) can be applied at frequencies of less than 1 kHz,1 Hz-900 Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250 Hz, and anyfrequencies therein. In some embodiments, nanoparticle formulationtreatments are performed alone, in combination, sequentially or repeated1-24 times. In other embodiments, the nanoparticles (e.g., photoactivenanoparticles, such as plasmonic nanoparticles) are capable ofselectively localizing to a first component of the skin, where physicalmassage or pressure, ultrasound, or heat increase the selectivelocalization of the nanoparticles to this first component. Additionally,the nanoparticles are selectively removable from components of the skinother than the first component, such removal accomplished with acetone,alcohol, water, air, peeling of the skin, chemical peeling, waxing, orreduction of the plasmonic compound. Further, in some embodiments thenanoparticles have a coat layer to increase solubility of thenanoparticles in the carrier and/or reduce “stickiness” and accumulationin non-target areas. The subject matter described herein also providesembodiments in which at least a portion of an exterior surface of thenanoparticle is modified, such as to include a layer of a polymer, polarmonomer, non-polar monomer, biologic compound, a metal (e.g., metallicthin film, metallic composite, metal oxide, or metallic salt), adielectric, or a semiconductor. Alternatively, the exterior surfacemodification is polar, non-polar, charged, ionic, basic, acidic,reactive, hydrophobic, hydrophilic, agonistic, or antagonistic. Incertain embodiments where at least one dimension of at least onenanoparticle within a solution of plasmonic nanoparticles is below50-100 nm, the nanoparticle surface can be coated with a matrix (e.g.silica) of 10-100 nm thickness or more in order to increase thatdimension or particle to 50-100 nm or more. This increased dimensionsize can increase the delivery of all nanoparticles to a target region(e.g., hair follicle) and limit delivery to non-target region (e.g.dermis).

Penetration Means

In some embodiments, the compositions of the instant disclosure aretopically administered. Provided herein are means to redistributeplasmonic particles and other compositions described herein from theskin surface to a component of dermal tissue including a hair follicle,a component of a hair follicle, a follicle infundibulum, a sebaceousgland, or a component of a sebaceous gland using high frequencyultrasound, low frequency ultrasound, massage, iontophoresis, highpressure air flow, high pressure liquid flow, vacuum, pre-treatment withFractionated Photothermolysis laser or derm-abrasion, or a combinationthereof. In several embodiments of the invention, low frequencyultrasound can be applied at frequencies of 1 kHz to 500 kHz, e.g., 1kHz-100 kHz, 5 kHz-45 kHz, 20 kHz-50 kHz, 30 kHz-40 kHz, 30 kHz, 40 kHz,and any ranges or frequencies therein.) In several embodiments of theinvention, massage (e.g., hand massage, vibration, mechanical vibration)can be applied at frequencies of less than 1 kHz, 1 Hz-900 Hz, 5-500 Hz,10-100 Hz, 1-80 Hz, 50-250 Hz, and any frequencies therein.

In some embodiments, a delivery device 200 is used to deliver,distribute, redeliver, redistribute, penetrate, drive, disperse, direct,and/or enhance movement of a composition 100 to a target location. Insome embodiments, the delivery device 200 is a mechanical vibrationdevice. In some embodiments, the delivery device 200 is a mechanicalvibration device configured for mechanical mixing. In one embodiment, amechanical vibration device vibrates at frequencies of less than 1 kHz,1 Hz-900 Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250 Hz, and anyfrequencies therein. In some embodiments a mechanical vibration devicevibrates, laterally, longitudinally, or radially. In one embodiment, amechanical vibration device vibrates longitudinally. In one embodiment,a mechanical vibration device vibrates horizontally. In one embodiment,a mechanical vibration device vibrates radially. In one embodiment, amechanical vibration device vibrates with a mix of one or more motions,longitudinal, horizontal, and/or radial. In one embodiment, a mechanicalvibration device vibrates longitudinally at 80 Hz. In one embodiment,the delivery device 200 is a mechanical vibration device with 80 Hzlongitudinal vibration. In one embodiment, the delivery device 200 is aVibraderm with 80 Hz longitudinal vibration, configured to deliver acomposition 100 to a depth of 1000 microns.

In some embodiments, the delivery device 200 is an ultrasound device. Insome embodiments, the delivery device 200 is an ultrasound device withfocused ultrasound energy. In some embodiments, the delivery device 200is an ultrasound device with unfocused ultrasound energy. In someembodiments, the delivery device 200 is an ultrasound device thatproduces cavitation. In some embodiments, the delivery device 200 is anultrasound device with pulsed energy. In some embodiments, the deliverydevice 200 is an ultrasound device with non-pulsed energy. In someembodiments, the delivery device 200 is an ultrasound device withsurface localized energy such as that which can be generated by aSonotrode. A sonotrode may refer to an acoustic horn, acousticwaveguide, ultrasonic probe, or ultrasonic horn. A sonotrode consists ofa metal shaft, rod, horn, cone, taper, barbell, wedge, or other shapecapable of translating and/or augment the amplitude produced by a lowfrequency ultrasonic transducer. The main function of a sonotrode is todeliver ultrasonic energy from a transducer into a gas, liquid, solid ortissue. In some embodiments, delivery device 200 is a sonotrode that isattached to an ultrasonic transducer or a stack of ultrasonictransducers. In some embodiments, the sonotrode delivery device 200 isdesigned to operate at frequencies between 15 kHz-100 kHz, e.g., 15kHz-85 kHz, 15 kHz-75 kHz, 15 kHz-65 kHz, 15 kHz-55 kHz, 15 kHz-45 kHz,15 kHz-35 kHz, 15 kHz-25 kHz, 25 kHz-60 kHz, 20 kHz-60 kHz, 20 kHz-50kHz, 20 kHz-40 kHz, and any frequencies therein. In some embodiments ofdelivery device 200, the sonotrode is used to deliver ultrasonic energyinto the composition, tissue, and/or surface of the tissue or anycombination of the composition, tissue, and surface of the tissue. Insome embodiments of delivery device 200, the power and amplitude of thesonotrode generates acoustic cavitation in the surrounding medium (e.g.,composition, tissue, and/or surface of the tissue or any combination ofthe composition, tissue, and surface of the tissue). In some embodimentsof the delivery device 200, the action of sonotrode generates heat,mixing, jetting, and streaming at or near the surface of the sonotrodein the surrounding medium. In some embodiments of delivery device 200,the acoustic waveguide within the sonotrode may also produce evanescentwaves responsible for localized effects in the surrounding medium. Oneor more of the actions of the sonotrode are responsible for driving thedelivery of the composition into the targeted area of the skin.Localized surface effects of the sonotrode include cavitation, jetting,mixing, and streaming.

In some embodiments, the delivery device 200 is a high frequencyultrasound device. In some embodiments, the delivery device 200 is a lowfrequency ultrasound device. In several embodiments of the invention,low frequency ultrasound can be applied at frequencies of 1 kHz to 500kHz, e.g., 1 kHz-100 kHz, 5 kHz-45 kHz, 20 kHz-50 kHz, 30 kHz-40 kHz, 30kHz, 40 kHz, and any ranges or frequencies therein.) In one embodiment,the delivery device 200 operates at a frequency of 20-50 kHz and morespecifically 32.4 kHz with an axial amplitude between 1 micron-30microns, e.g., 1 micron-20 microns, 1 micron-15 microns, 5 microns-15microns, 5 microns-12 microns, 7 microns, 8 microns, 9 microns, 10microns, 11 microns, 12 microns,-15 microns and any range of axialamplitudes therein. In one embodiment, the delivery device 200 operatesat a frequency of 32.4 kHz with an axial amplitude between 1 micron-20microns and a radial amplitude between 0 microns-5 microns, e.g., 0microns, 1 micron, 2 microns, 3 microns, 4 microns, 5 microns, and anyradial amplitude therein. In one embodiment, the delivery device 200operates at a frequency of 32.4 kHz with an axial amplitude between 1micron-20 microns, radial amplitude between 0 microns-5 microns, and isdriven by an ultrasonic transducer with powers between 3 W-20 W, e.g.,with surface localized energy configured to induce cavitation,configured to deliver a composition 100 to a depth of at least 1500microns.

In one embodiment, the delivery device 200 is a low frequency ultrasounddevice that operates at a frequency of 30-60 kHz non-pulsed ultrasoundconfigured to induce cavitation. In one embodiment, the delivery device200 is a low frequency ultrasound device that operates at a frequency of30-60 kHz non-pulsed ultrasound configured to induce cavitation from anunfocused transducer. In one embodiment, the delivery device 200 is alow frequency ultrasound device that operates at a frequency of 36 kHznon-pulsed ultrasound with a second harmonic at 55 kHz configured toinduce cavitation at depth of approximately 8 mm and 18 mm in water fromthe face of the unfocused transducer, operating at a power between 2W-15 W, and configured to deliver a composition 100 to a depth of atleast 1000 microns.

In some embodiments, the delivery device 200 is configured to deliver acomposition 100 to a depth of 1-100 microns, 1-1000 microns, 1-1500microns, 1-2000 microns, 1-3000 microns, 1-4000 microns, and/or 1-5000microns. In some embodiments, the delivery device 200 is configured todeliver a composition 100 to a depth of 1000-1500 microns. In someembodiments, the delivery device 200 is configured to deliver acomposition 100 to a depth of 1000 microns. In some embodiments, thedelivery device 200 is configured to deliver a composition 100 to adepth of 1500 microns. In some embodiments, the delivery device 200 isconfigured to deliver a composition 100 to a depth of 2000 microns. Insome embodiments, the delivery device 200 is configured to deliver acomposition 100 to a depth of 2500 microns. In some embodiments, thenanoparticles described herein are formulated to penetrate muchdeeper—up to several centimeters, or into the panniculus adiposus(hypodermis) layer of subcutaneous tissue. For example, the compositionscan be administered by use of a sponge applicator, cloth applicator,spray, aerosol, vacuum suction, high pressure air flow, high pressureliquid flow direct contact by hand ultrasound and other sonic forces,mechanical vibrations, physical manipulation, hair shaft manipulation(including pulling, massaging), physical force, thermal manipulation, orother treatments. Nanoparticle formulation treatments are performedalone, in combination, sequentially or repeated 1-24 times.

Specific Types of Applications

Non-limiting descriptions of certain applications are provided below.The nanoparticles described may be plasmonic or non-plasmonic. Forexample, non-plasmonic, photoactive nanoparticles may be used.

Acne Treatment

Acne is caused by a combination of diet, hormonal imbalance, bacterialinfection (Propionibacterium acnes), genetic predisposition, and otherfactors. The nanoparticle-based methods and systems described herein foracne treatment are able to focally target causative regions of thedermis, the sebaceous gland and the hair follicle, and thus haveadvantages compared to the existing techniques known in the art,including chemical treatment (peroxides, hormones, antibiotics,retinoids, and anti-inflammatory compounds), dermabrasion, phototherapy(lasers, blue and red light treatment, or photodynamic treatment), orsurgical procedures.

In particular, laser-based techniques are becoming an increasinglypopular acne treatment, but a substantial limitation is the lack ofselective absorptive properties among natural pigments (e.g. fat, sebum)for specific wavelengths of light such that selective heating of onecell, structure, or component of tissue, particularly in the sebaceousglands, infundibulum, and regions of the hair follicle, is not achievedwithout heating of adjacent off-target tissue. The nanoparticlesdescribed herein provide significantly higher photothermal conversionthan natural pigments enabling laser energy to be focused to specificcells, structures, or components of tissue within the sebaceous gland,infundibulum, or regions of the hair follicle for selective photothermaldamage.

Using the materials and techniques described herein may provide acnetreatments of greater duration than existing methodologies. In certainembodiments, tuned selective ablation of the sebaceous gland orinfundibulum is achieved as described herein. In particular, plasmonicnanoparticles are specifically localized to regions of hair follicles inor proximate to the sebaceous gland or infundibulum.

Plasmonic nanoparticles exhibit strong absorption at wavelengths emittedby standard laser hair removal devices (e.g., 755 nm, 810 nm, 1064 nm)and intense pulse light (IPL) devices (e.g., 515-1200 nm range) relativeto surrounding epidermal tissue. Thus, irradiation of targeted plasmonicnanoparticles with laser light or IPL induces heat radiation from theparticles to the adjacent sebum, sebaceous gland, infundibulum, andother acne causing agents.

Hair Removal

The nanoparticle-based methods and systems described herein for skintreatment have advantages compared to the existing techniques known inthe art, including laser-based techniques, chemical techniques,electrolysis, electromagnetic wave techniques, and mechanical techniques(e.g., waxing, tweezers). Such techniques fail to adequately providepermanent hair removal across a breadth of subjects. In particular,subjects having light to medium-pigmented hair are not adequately servedby these techniques, which suffer from side-effects including pain andthe lack of beneficial cosmetic affects including hair removal.Laser-based techniques are popular in a variety of applications, but asubstantial limitation is the lack of selective absorptive propertiesamong natural pigments (e.g. melanin) for specific wavelengths of lightsuch that selective heating of one cell, structure, or component oftissue is achieved without heating of adjacent off-target tissues. Thenanoparticles described herein provide significantly higher photothermalconversion than natural pigments enabling laser energy to be focused tospecific cells, structures, or components of tissue for selectivephotothermal damage. The methods described herein are useful for hairremoval of all types and pigmentations. For example, melanin, thepredominant hair pigment, is an aggregation of chemical moietiesincluding eumelanin and phaeomelanin. Eumelanin colors hair grey, black,yellow, and brown. A small amount of black eumelanin in the absence ofother pigments causes grey hair. Types of eumelanin include blackeumelanin and brown eumelanin, with black melanin being darker thanbrown. Generally, black eumelanin predominates in non-European subjectsand aged Europeans, while brown eumelanin is in greater abundance inyoung European subjects. Phaeomelanin predominates in red hair. Inanother example, vellus hair (“peach fuzz”) is a type of short, fine,light-colored, and usually barely noticeable hair that develops on muchor most of a subject's body (excluding lips, palms of hand, sole offoot, navel and scar tissue). While the density of vellus hair isgenerally lower than that of other hair types, there is variation fromperson to person in the density, thickness, and pigmentation. Vellushair is usually less than 2 mm long and the follicle containing thevellus hair is generally not connected to a sebaceous gland. Conditionsassociated with an overabundance of vellus hair include Cushing'ssyndrome and anorexia nervosa, such overgrowth being treatable using themethods and compositions described herein. Further, provided are methodsof targeting hair growth at a given stage. Hair grows in cycles ofvarious stages or phases. Growth phase is termed “anagen”, while“catagen” includes the involuting or regressing phase, and “telogen”encompasses the resting or quiescent phase. Each phase has severalmorphologically and histologically distinguishable sub-phases.Generally, up to 90% of the hair follicles on a subject are in anagenphase (10-14% are in telogen and 1-2% in catagen). The cycle's length isgoverned by cytokines and hormones, and varies on different parts of thebody. For eyebrows, the cycle is completed in around 4 months, while ittakes the scalp 3-4 years to finish. The methods and compositionsdescribed herein are sufficient to treat hair of all growth stages orphases.

More permanent reduction or removal of all hair types is providedherein, relative to hair removal treatments known in the art. In certainembodiments, tuned selective ablation of the hair shaft and destructionof stem cells in the bulge region is provided, as described herein. Inparticular, plasmonic nanoparticles are specifically localized toregions of hair follicles in or proximate to the bulge region, a stemcell-rich domain of the hair follicle. Moreover, the plasmonicnanoparticles are localized in close approximation of ˜50-75% of thehair shaft structure.

Plasmonic nanoparticles exhibit strong absorption at wavelengths emittedby standard laser hair removal devices (e.g., 755 nm, 810 nm, 1064 nm)and intense pulse light (IPL) devices (e.g., 515-1200 nm range) relativeto surrounding epidermal tissue. Thus, irradiation of targeted plasmonicnanoparticles with laser light induces heat radiation from the particlesto the adjacent stem cells (or in some cases, the architecture of thehair shaft itself), resulting in cell death and a disruption of thenormal regenerative pathway.

Non-malignant and Malignant Skin Tumors

Laser therapies for the prevention and treatment of non-malignant,malignant, melanoma and non-melanoma skin cancers have been focusedlargely on photodynamic therapy approaches, whereby photosensitiveporphyrins are applied to skin and used to localize laser light, producereactive oxygen species and destroy cancer cells via toxic radicals. Forexample, 5-ALA combined with laser treatment has been FDA-approved forthe treatment of non-melanoma skin cancer actinic keratoses, and it isused off-label for the treatment of widely disseminated, surgicallyuntreatable, or recurrent basal cell carcinomas (BCC). However, thisprocedure causes patients to experiences photosensitivity, burning,peeling, scarring, hypo- and hyper-pigmentation and other side effectsdue to non-specific transdermal uptake of porphyrin molecules. Thenanoparticles described herein provide significantly higher photothermalconversion than natural pigments and dyes, enabling laser energy to befocused to specific cells, structures, or components of tissue forselective thermomodulation

Using the materials and techniques described herein may provide cancertreatments of greater degree and duration than existing methodologies.In certain embodiments, tuned selective ablation of specific targetcells, such as Merkel cells or Langerhans cells, as described herein. Inparticular, plasmonic nanoparticles are specifically localized toregions of hair follicles where follicular bulge stem cells arise toform nodular basal cell carcinomas and other carcinomas. Plasmonicnanoparticles may also be delivered to other target cells that causetumors, for example, the interfollicular epithelium, which include thecell of origin for superficial basal cell carcinomas.

Plasmonic nanoparticles exhibit strong absorption at wavelengths emittedby standard laser hair removal devices (e.g., 755 nm, 810 nm, 1064 nm)and intense pulse light (IPL) devices (e.g., 515-1200 nm range) relativeto surrounding epidermal tissue. Thus, irradiation of targeted plasmonicnanoparticles with laser light induces heat radiation from the particlesto the adjacent keratinocyte, melanocyte, follicular bulge stem cell,cancer cell, or cancer cell precursor, resulting in cell death orinhibited cell growth for cancer prevention and treatment.

Subdermal Applications. Target tissues for subdermal applicationsinclude the adipose tissue and connective tissue below the integumentarysystem. Diseases or conditions suitable for treatment withsubdermatological applications include wrinkles and tattoos. Otherapplications include skin rejuvenation and/or resurfacing, the removalor reduction of stretch marks and fat ablation.

Vascular Applications. Target tissues for vascular applications includearteries, arterioles, capillaries, vascular endothelial cells, vascularsmooth muscle cells, veins, and venules. Diseases or conditions suitablefor treatment with vascular applications include spider veins, leakyvalves, and vascular stenosis. In particular, vein abnormalities accountfor a substantial proportion of cosmetic diseases or conditionsaffecting the vasculature. Individuals with vein abnormalities such asspider veins or faulty venous valves suffer from pain, itchiness, orundesirable aesthetics.

Additionally, there are several indication for which ablation of othervessels including arteries, arterioles, or capillaries could providetherapeutic or cosmetic benefit including: 1) ablation of vasculaturesupplying fat pads and/or fat cells, 2) ablation of vasculaturesupporting tumors/cancer cells, 3) ablation of vascular birth marks(port-wine stains, hemangiomas, macular stains), and 4) any otherindication whereby ablation of vessels mediates the destruction oftissue and apoptosis or necrosis of cells supported by those vesselswith therapeutic or cosmetic benefit. Provided herein are methods forusing the compositions described herein for the selective destruction ofcomponent(s) of veins from plasmonic nanoparticles focally or diffuselydistributed in the blood. Plasmonic nanoparticles are combined with apharmaceutically acceptable carrier as described above and areintroduced into the body via intravenous injection. Nanoparticlesdiffuse into the blood and, in some embodiments, localize to specificvascular tissues. Subsequently, the nanoparticles are activated withlaser or light-based systems as known in the art for treating skinconditions such as hair removal or spider vein ablation. Alternatively,image or non-image guided fiber optic waveguide-based laser or lightsystems may be used to ablate vessel or blood components in largerveins. In one embodiment, a device with dual functions for bothinjecting nanoparticles and administering light through on opticalwaveguide may be used. Activated nanoparticles heat blood and adjacenttissue (vessels, vessel walls, endothelial cells, components on or inendothelial cells, components comprising endothelial basement membrane,supporting mesenchymal tissues, cells, or cell components around thevessel, blood cells, blood cell components, other blood components) toablative temperatures (38-50 degrees C. or higher).

Provided herein is a composition comprising a pharmaceuticallyacceptable carrier and a plurality of plasmonic nanoparticles in anamount effective to induce thermomodulation of a vascular orintravascular target tissue region with which the composition isintravenously contacted. Furthermore, the composition of plasmonicnanoparticle may comprise a microvascular targeting means selected fromthe group consisting of anti-microvascular endothelial cell antibodiesand ligands for microvascular endothelial cell surface receptors. Alsoprovided is a method for performing thermoablation of a target vasculartissue in a mammalian subject, comprising the steps of contacting aregion of the target vascular tissue with a composition comprising aplurality of plasmonic nanoparticles and a pharmaceutically acceptablecarrier under conditions such that an effective amount of the plasmonicnanoparticles localize to a domain of the target vascular region; andexposing the target tissue region to energy delivered from a surfaceplasmon resonance excitation sources (e.g., nonlinear excitation surfaceplasmon resonance source) in an amount effective to inducethermoablation of the domain of the target vascular region.

Oral and nasal applications. Target tissues for oral applicationsinclude the mouth, nose, pharynx, larynx, and trachea. Diseases orconditions suitable for treatment with vascular applications includeoral cancer, polyps, throat cancer, nasal cancer, and Mounier-Kuhnsyndrome. Other conditions suitable for treatment include allergies orvoice disorders involving vocal chords.

Endoscopic Applications. Target tissues for endoscopic applicationsinclude the stomach, small intestine, large intestine, rectum and anus.Diseases or conditions suitable for treatment with vascular applicationsinclude gastrointestinal cancer, ulcerative colitis, Crohn's disease,Irritable Bowel Syndrome, Celiac Disease, Short Bowel Sydrome, or aninfectious disease such as giardiasis, tropical sprue, tapeworminfection, ascariasis, enteritis, ulcers, Whipple's disease, andmegacolon.

Methods of thermomodulation. Provided are embodiments of methods forperforming thermomodulation of a target tissue region. A nanoparticlecomposition comprising a plurality of plasmonic nanoparticles underconditions such that an effective amount of the plasmonic nanoparticleslocalize to a domain of the target tissue region; and exposing thetarget tissue region to energy delivered from a surface plasmonresonance excitation sources (e.g., nonlinear excitation surface plasmonresonance source) in an amount effective to induce thermomodulation ofthe domain of the target tissue region.

Removal of non-specifically bound nanoparticles. In various embodiments,removing nanoparticles localized on the surface of the skin may beperformed by contacting the skin with acetone, alcohol, water, air, adebriding agent, or wax. Alternatively, physical debridement may beperformed. Alternatively, one can perform a reduction of the plasmonicor other compound.

Amount of energy provided. In some embodiments, skin is irradiated at afluence of 1-60 Joules per cm² with laser wavelengths of about, e.g.,750 nm, 810 nm, 1064 nm, or other wavelengths, particularly in the rangeof infrared light. Various repetition rates are used from continuous topulsed, e.g., at 1-10 Hz, 10-100 Hz, 100-1000 Hz. While some energy isreflected, it is an advantage of the subject matter described herein isthat a substantial amount of energy is absorbed by particles, with alesser amount absorbed by skin. Nanoparticles are delivered to the hairfollicle, infundibulum, or sebaceous gland at concentration sufficientto absorb, e.g., 1.1-100× more energy than other components of the skinof similar volume. This is achieved in some embodiments by having aconcentration of particles in the hair follicle with absorbance at thelaser peak of 1.1-100× relative to other skin components of similarvolume.

To enable tunable destruction of target skin structures (e.g., sebaceousglands, infundibulum, hair follicles), some embodiments oflight-absorbing nanoparticles are utilized in conjunction with a laseror other excitation source of the appropriate wavelength. The laserlight may be applied continuously or in pulses with a single or multiplepulses of light. The intensity of heating and distance over whichphotothermal damage will occur are controlled by the intensity andduration of light exposure. In some embodiments, pulsed lasers areutilized in order to provide localized thermal destruction. In some suchembodiments, pulses of varying durations are provided to localizethermal damage regions to within 0.05, 0.1, 0.5, 1, 2, 5, 10, 20, 30,50, 75, 100, 200, 300, 500, 1000 microns of the particles. Pulses are atleast femtoseconds, picoseconds, microseconds, or milliseconds induration. In some embodiments, the peak temperature realized in tissuefrom nanoparticle heating is at least 5, 10, 15, 20, 25, 30, 40, 50, 60,70, 80, 90, 100, 200, 300, or 500 degrees Celsius. In some embodimentsthat utilize pulsed heating, high peak temperatures are realized locallywithin the hair shaft without raising the macroscopic tissue temperaturemore than 0.1, 0.5, 1, 2, 3, 4, 5, 7, 9, 12, 15, or 20 degrees Celsius.In some embodiments short pulses (100 nanoseconds-1000 microseconds) areused to drive very high transient heat gradients in and around thetarget skin structure (e.g., sebaceous gland and/or hair follicle) fromembedded particles to localize damage in close proximity to particlelocation. In other embodiments, longer pulse lengths (1-10 ms, or 1-500ms) are used to drive heat gradients further from the target structureto localize thermal energy to stem cells in the bulge region or othercomponents greater than 100 μm away from the localized particles.Fluences of 1-10 Joules per cm² or 1-30 Joules per cm² are generallysufficient to thermally ablate follicles that have high particleconcentrations and thus higher absorbance than skin (e.g., 1.1-100 timesper volume absorbance of skin). These fluences are often lower than whatis currently employed (e.g., Diode: 25-40 J/cm², Alexandrite: 20 J/cm2,Nd:YAG: 30-60 J/cm²) and lead to less damage to non-follicular regions,and potentially less pain.

Plasmon Resonance Systems. Provided are embodiments of plasmon resonancesystems containing a surface that includes a plurality of plasmonicnanoparticles, and a nonlinear excitation source. Optionally, the systemcontains a means to generate thermal heating of the surface. Preferably,the surface is a component of skin that is targeted for cosmetic ortherapeutic treatment (e.g., bulge region for hair removal, infundibulumor sebaceous gland for acne prevention). Also provided as a component ofthe system is a means for delivering plasmonic nanoparticles to the skinsurface, such as an applicator, a spray, an aerosol, vacuum suction,high pressure air flow, or high pressure liquid flow. Further providedare means of localizing plasmonic nanoparticles to a component of theskin (e.g., hair follicle, bulge region, sebaceous gland, infundibulum).Useful surface delivery means include a device that generates highfrequency ultrasound, low frequency ultrasound, heat, massage, contactpressure, or a combination thereof. In several embodiments of theinvention, low frequency ultrasound can be applied at frequencies of 1kHz to 500 kHz, e.g., 1 kHz-100 kHz, 5 kHz-45 kHz, 20 kHz-50 kHz, 30kHz-40 kHz, 30 kHz, 40 kHz, and any ranges or frequencies therein.) Inseveral embodiments of the invention, massage (e.g., hand massage,vibration, mechanical vibration) can be applied at frequencies of lessthan 1 kHz, 1 Hz-900 Hz, 5-500 Hz, 10-100 Hz, 1-80 Hz, 50-250 Hz, andany frequencies therein.

Further provided are systems that contain a removal means for removingnanoparticles on a non-follicular portion of the skin. The removal meansincludes at least one of acetone, alcohol, water, air, chemical peeling,wax, or a compound that reduces the plasmonic compound.

In addition, the systems of the present disclosure provide nonlinearexcitation source that generates a continuous wave optical source or apulsed optical source. Alternatively, the nonlinear excitation source iscapable of generating electromagnetic radiation, ultrasound, thermalenergy, electrical energy, magnetic energy, or electrostatic energy.Provided are systems wherein the nonlinear excitation source is capableof irradiating the nanoparticles with an intensity from about 0.00005mW/cm² to about 1000 TW/cm². Further, the nonlinear excitation source iscapable of functioning in a one-photon mode, two-photon mode,multi-photon mode, step-wise mode, or up-conversion mode. A fiber, awaveguide, a contact tip, or a combination thereof may be used in theinstant systems.

In some embodiments, the system contains a monitoring device such as atemperature sensor or a thermal energy detector. In other embodiments,the systems also contain a controller means for modulating the nonlinearexcitation source (e.g., a “feedback loop controller”). In a relatedembodiment, the system contains a means for detecting a temperature ofthe surface or a target tissue adjacent to the surface, wherein thecontroller means modulates the intensity of the nonlinear excitationsource and/or the duration of the excitation. In such embodiments, thecontroller means preferably modulates the intensity of the nonlinearexcitation source such that a first component of the hair follicle isselectively thermoablated relative to a second component of the hairfollicle. In further embodiments, a cooling device is directly contactedwith the skin during irradiation to minimize the heating ofnanoparticles or skin at the surface, while nanoparticles that havepenetrate more deeply into the follicle, skin, or sebaceous gland heatto temperatures that selectively ablate the adjacent tissues.

Skin is one embodiment of a target tissue. The skin preferably containsa hair follicle and/or a sebaceous gland, where the nonlinear excitationsource generates energy that results in heating the skin in an amounteffective to induce thermomodulation of a hair follicle, a infundibulum,a sebaceous gland, or a component thereof, such as by heating sufficientto cause the temperature of the skin to exceed 37° C., such as 38° C.,39° C., 40° C., 41° C., 42° C., 43° C., 44° C., 45° C., 46° C., 47° C.,48° C., 49° C., to about 50° C. or greater.

Methods of Formulation. In some embodiments, methods for formulating thenanoparticles of the present disclosure into a form suitable for use aredescribed herein. In one embodiment, the nanoparticle compositions aregenerated by:

-   -   a) forming a first mixture containing a plurality of        nanoparticles and a first solvent;    -   b) exchanging the first solvent for a second solvent to form a        second mixture; and    -   c) combining the second mixture and a cosmetically or        pharmaceutically acceptable carrier; thereby forming a        nanoparticle composition.

The exchanging step is optionally performed using liquid chromatography,a solvent exchange system, a centrifuge, precipitation, or dialysis.Preferably, the nanoparticles are surface modified through a controlledreduction step or an oxidation step. Such surface modification mayinvolve a coating step, such as the adsorbance of a monomer, polymer, orbiological entity to a surface of the nanoparticle. Typically, thecoating step involves contacting the nanoparticles with an oxidativeenvironment. Further, the coating step may include monomerpolymerization to create polymer coat.

In one embodiment, the methods described herein may also include thesteps of dissolving the nanoparticles in a non-polar solvent andsubsequently mixing the dissolved nanoparticles with a polar solvent soas to encapsulate the nanoparticles in an emulsion. Further, theaddition of surfactants (e.g. sodium dodecyl sulfate, sodium laureth2-sulfate, ammonium lauryl sulfate, sodium octech-1/deceth-1 sulfate) atconcentrations of 0.1-10% may be used to disrupt the epidermal skinbarrier, emulsify the sebum and enable improved mixing of hydrophilicnanoparticles in aqueous solutions. Further, a concentration of thenanoparticles such as centrifugation or lyophilization may be employed.Further, the nanoparticles may be pretreated with heat or radiation.Also provided is the optional step of conjugating a biological entity orplurality of biological entities to the nanoparticles. Such aconjugating step may involve a thiol, amine, or carboxyl linkage of thebiological entities to the nanoparticles.

Diseases and Disorders. Several embodiments of the present disclosurecan be used on human (or other animal) skin for the treatment ofwrinkles and other changes related to photo-aging or chronologic aging(generally termed skin rejuvenation), for the treatment of diseasesincluding skin diseases, for the reduction of acne and related disorderssuch as rosacea, folliculitis, pseudofolliculitis barbae orproliferative or papulosquamous disorders such as psoriasis, for thestimulation or reduction of hair growth, and for reduction of cellulite,warts, hypopigmentation such as port-wine stain (PWS; nevus flammeus),birthmarks, hyperhidrosis, varicose veins, pigment problems, tattoos,vitiligo, melasma, scars, stretch marks, fungal infections, bacterialinfections, dermatological inflammatory disorders, musculoskeletalproblems (for example, tendonitis or arthritis), to improve healing ofsurgical wounds, burn therapy to improve healing and/or reduce andminimize scarring, improving circulation within the skin, and the like.

Several embodiments of the present disclosure can also be useful inimproving wound healing, including but not limited to chronic skinulcers, diabetic ulcers, gastric ulcers, thermal burn injuries, viralulcers or disorders, periodontal disease and other dental disease. Thepresent disclosure can be useful in treating the pancreas in diabetes.The present disclosure can be useful for in vitro fertilizationenhancement, and the like. The present disclosure, in certainembodiments, is also useful in enhancing the effects of devices thatcreate an injury or wound in the process of performing cosmetic surgeryincluding non-ablative thermal wounding techniques for treating skinwrinkles, scars, stretch marks and other skin disorders. Under suchcircumstances, it may be preferable to use conventional non-ablativethermal treatments in combination with the methods of the presentdisclosure. The instant application, in certain embodiments, are used inconjunction with micro- or surface abrasion, dermabrasion, or enzymaticor chemical peeling of the skin or topical cosmeceutical applications,with or without nanoparticle application to enhance treatment, as theremoval of the stratum corneum (and possibly additional epitheliallayers) can prove beneficial for some treatment regimen. The methods ofthe present disclosure are particularly applicable to, but are notlimited to, acne treatment, hair removal, hair growth/hair folliclestimulation, reduction/prevention of malignant and non-malignant skintumors, and skin rejuvenation, as described herein.

The dermatologically therapeutic methods described herein may be formedusing nanoparticle irradiation alone, nanoparticle irradiation incombination with nano- or microparticles, or nanoparticle irradiationwith a composition comprising nano- or microparticles and one or moretherapeutic agents. Such nanoparticle irradiation may be produced by anyknown nanoparticle generator, and is preferably a focused nanoparticlegenerator capable of generating and irradiating focused nanoparticlewaves. Additionally, nanoparticle waves can be focused in tissues toprovide damage to local areas with a desirable size and shape.

Several embodiments of the invention describe the development andutilization of compositions containing photoactive materials (e.g.,nanoparticles and other materials) for the treatment of small targetregions of skin including acne scars and other skin conditions. In someembodiments, such compositions are generally applied topically, throughan apparatus that provides the composition in a form suitable forcontact with and retention at a target region of skin in a manner thatencompasses irradiating the skin with light (e.g., electromagneticradiation) having a wavelength sufficient to ablate or otherwise damagethe target region of skin and cause remodeling of the skin tissue.Without being bound by theory, it is believed that the damage to theskin resulting from heat transfer from the photoactive material afterinteraction with the radiation induces a wound healing response,including new extracellular matrix (e.g., collagen) production andremodeling, neovascularization, and epidermal normalization. As providedherein, “thermal injury” encompasses cell death in one or more regionsof the dermal tissue of interest (“lethal damage”), or stimulation ofthe release of cytokines, heat shock proteins, and other wound healingfactors without stimulating necrotic cell death (“sublethal damage”).

In one embodiment, provided are methods for reducing dermal scar tissue,typically in order to improve the appearance of the skin tissuecontaining the scar, in a human subject using photoactive materials.While humans are provided as one example of mammalian subjects, one ofskill in the art would recognize that other mammals are suitable fortreatment herewith.

Typical dermal scar tissue can result from acne infection or other acuteor chronic damage or injuries, such as burns, puncture or abrasiveinjury, surgery, or from conditions caused by environmental conditionsor inherited genetic aberrations.

While scars are three dimensional in nature, description of theepidermal surface of the scar on the surrounding skin tissue can beaccomplished by provision of a depth, length and a width of the scar,each of which may be, e.g., less than about 1 mm, or about 2, 3, 4, 5,6, 7, 8, 9, 10 mm or greater than 10 mm, or provision of the surfacearea encompassed by the scar, e.g., under about 5 mm², such as about 10mm², 15 mm², 20 mm², 25 mm², 30 mm², 35 mm², 40 mm², 45 mm², 50 mm², orgreater than about 50 mm². Dermal scars generally have a non-uniformdepth, and are generally described as extending from the epidermal layerinto the dermis, and optionally through the dermis. Typically, an acnescar on the chin, cheek, forehead or other facial area is at least 0.01mm, 0.25 mm, 0.50 mm, 1 mm, 2 mm, 3 mm, 4 mm, or 5 mm in medianthickness, or any thickness in the range of 0.01 mm to 5 mm.

Skin tissue. In preferred embodiments, a region of skin tissue on ahuman subject, herein a “target region”, is subjected to the methodsprovided herein. Optionally, more than one target region is treatedduring a treatment regimen, and such treatment regimens may happen once,or more than once, e.g., once or several times per month, once orseveral times per week, once or several times per day, within hours orwithin less than one hour. A target region contains an epidermalsurface, which contains the skin feature to be treated, such as an acnescar or other dermal scar tissue (also termed a “lesion” herein). Thescar tissue may be newly present (e.g., within days, weeks, or a fewmonths of formation following the damage or injury) or may be of longerduration (e.g., several months, or years). In some embodiments, a “smalltarget region” is treated, meaning a dermal scar region that does notexceed 25 mm², and or does not have a longest surface dimension greaterthan 5 mm; such small target regions are generally located on the faceor neck after acne vulgaris infection, or one of many inflammatorydiseases such as chicken pox or small pox. Small target regions ofdermal scars may be atrophic, hypertrophic, keloidal, or may lay largelywithin the planar surface of the skin, but have irregular texture,contour, or edges, in various embodiments, small target regions ofdermal scars are termed rolling scars, ice-pick scars, and box-carscars.

Photoactive materials. Photoactive materials, particles or nanoparticles(also termed “photoresponsive” materials and “photoabsorbable”materials) include chromophores and plasmonic nanoparticles. Achromophore is able to selectively absorb a chosen wavelength of lightthereby enhancing the effectiveness of the irradiation, such as laserlight.

In one embodiment, photoactive materials include plasmonic nanoparticleswith a nanoparticular metallic structure within which localized surfaceplasmons are excited by light. These surface plasmons are surfaceelectromagnetic waves that propagate in a direction parallel to themetal/dielectric interface (e.g., metal/air or metal/water). Atresonance wavelengths plasmonic nanoparticles are non-linear absorbersof energy, whereby both incident light energy and energy from light notdirectly incident on the particle is coupled to and absorbed by theparticle. In various embodiments, nanoparticle compositions, andformulations containing nanoparticle compositions, contain from about10⁹ to about 10¹⁶ nanoparticles per ml, such as 10⁹, 10¹⁰, 10¹¹, 10¹²,10¹³, 10¹⁴, 10¹⁵, or 10¹⁶ particles. In one embodiment, the compositionscontain a sufficient concentration of particles so that the amount ofparticles localized to an effective 0.01-0.05 ml treatment volumes isfrom 10⁷ to 10¹³. In certain embodiments wherein increased concentrationof nanoparticles to a target region is desired, compositions containparticle concentrations with optical densities (O.D.) of 10 O.D.-1,000O.D., or optical densities greater than 1,000 O.D. In some embodimentsthese correspond to concentrations of about 0.01-10% w/w or more ofnanoparticles.

Nanoparticles may be homogenous or heterogeneous in size and othercharacteristics. In certain embodiments where optimal plasmonicresonance is desired, a particle size in the range of from about 10 nmto about 200 nm is provided. Alternatively, in embodiments whereenhanced penetration of the nanoparticles into a target tissue regionsuch as a scar is desired, a particle size in the range of from about200 nm to about 1000 nm is provided. Modulation of particle size presentin the composition is also a useful means of concentrating thecomposition in a target domain. Further, as described herein,nanoparticles having a size range of from about 10 nm to about 100 nmcan be used as component of a larger molecular structure, generally inthe range of from about 100 nm to about 1000 nm or more. For example,photoactive particles, such as plasmonic nanoparticles, can be surfacecoated to increase its size, embedded into an acceptable carrier, or itcan be cross-linked or aggregated to other particles, or to othermaterials, that generate a larger particle. In certain embodiments whereat least one dimension of at least one nanoparticle within a solution ofplasmonic nanoparticles is below 50-100 nm, the nanoparticle surface canbe coated with a matrix (e.g. silica) of 10-100 nm thickness or more inorder to increase that dimension or particle to 50-100 nm or more. Thisincreased dimension size can increase the delivery of all nanoparticlesto a target region (e.g., scar tissue) and limit delivery to non-targetregion (e.g. surrounding non-scar epidermis).

Composition of particles. In various embodiments, the photoactiveparticle (e.g., nanoparticle) is a metal (e.g., gold, silver), metalliccomposite (e.g., silver and silica, gold and silica), metal oxide (e.g.iron oxide, titanium oxide), metallic salt (e.g., potassium oxalate,strontium chloride), intermetallic (e.g., titanium aluminide, alnico),electric conductor (e.g., copper, aluminum), electric superconductor(e.g., yttrium barium copper oxide, bismuth strontium calcium copperoxide), electric semiconductor (e.g., silicon, germanium), dielectric(e.g., silica, plastic), and/or a quantum dot (e.g., zinc sulfide,cadmium selenium). In non-limiting examples, the materials are gold,silver, nickel, copper, platinum, titanium, palladium, silicon,galadium, including any alloys, composites, and amalgams of thesemetals. Alternatively, the nanoparticle contains a composite including ametal and a dielectric, a metal and a semiconductor, or a metal,semiconductor and dielectric.

In one embodiment, the composition contains coated particles (e.g.,nanoparticles). Such coatings include a biorecognitive such as anantibody, a bioactive moiety such as a protein, or a biological materialthat is sourced from living matter. In one embodiment the compositioncontains an insulator such as silicon, or a thin metal coating such asgold, silver, nickel, platinum, titanium, or palladium. The compositionmay contain a peptide, a nucleic acid, a protein, or an antibody, or maycontain charged moieties, whereby those charges mediate enhanced ordiminished binding to the target skin.

Optical absorption. In several embodiments, particles have opticalabsorption qualities of about 10 nm to about 10,000 nm, e.g., 200-700nm, 700-1200 nm. In specific embodiments, the particles have opticalabsorption useful to excitation by standard laser devices or other lightsources. For example, in some embodiments, particles absorb atwavelengths of about 755 nm (alexandrite lasers), in the range of about800-810 nm (diode lasers), or about 1064 nm (Nd: YAG lasers). Similarly,in some embodiments, the particles absorb intense pulsed light (IPL),e.g., at a range of about 500 nm to about 1200 nm

Other chromophores can be useful in the present invention. The term“chromophore” shall be given its ordinary meaning and shall also includecompounds having chromophoric groups such as nitro groups, azo, alkyleneunits, esters, carbonyl groups, aldehydes, alkynes, aromatic rings,heterocyclics, carboxylic acids and the like. Photoactive materialsfunction as therapeutic or cytotoxic agents upon irradiation but aresubstantially inert prior to irradiation. A photoactive material may bea substance (solid, liquid, or gas) that has color or imparts a color tothe intact nanoparticles or microparticles (including when the substanceitself lacks color, for example, a clear gas, but scatterselectromagnetic waves, for example, light, and thus may appear colored,for example, white, blue, green, or yellow, depending on its scatteringproperties) under some conditions, for example, all of the time or afterexposure to a certain wavelength (such as in a fluorescent substance).For example, a chromophore can be a fluorescent, phosphorescent,wavelength up-converting, or other substance that may normally besubstantially invisible, but that emits ultraviolet, visible, orinfrared wavelengths during and/or after exposure to wavelengths from aparticular region of the electromagnetic spectrum. A chromophore canalso be a substance that reversibly or irreversibly changes colorspontaneously or in response to any stimulus. The chromophore can be orinclude rifampin, β-carotene, tetracycline, indocyanine green, Indiaink, Evan's blue, methylene blue, FD&C Blue No. 1 (Brilliant Blue FCF),FD&C Green No. 3 (Fast Green FCF), FD&C Red No. 3 (Erythrosine), FD&CRed No. 40, FD&C Yellow No. 5 (Tartrazine), or FD&C Yellow No. 6 (SunsetYellow FCF). The chromophore can be any colored substance approved bythe United States Food and Drug Administration for use in humans. Incertain embodiments, the chromophore can be detected by the naked eyeunder normal lighting conditions or when exposed to UV, near-UV, IR, ornear-IR radiation.

In various embodiments, photoactive particle (e.g., chromophores) may beprovided as a microparticle or a nanoparticle. As used herein, amicroparticle may be a particle of a relatively small size, notnecessarily in the micron size range; the term is used in reference toparticles of sizes that can be implanted to form tissue markings andthus can be less than 50 nm to 100 microns or greater. A micro- ornanoparticle may be of composite construction and is not necessarily apure substance; it may be spherical or any other shape. Microparticlesinclude, but are not limited to, (i) an indispersible, biologicallyinert coating, (ii) a core enveloped within the coating, wherein thecore includes the chromophore which is detectable through the coatingand is dispersible in the tissue upon release from the microparticle,and, optionally, (iii) an absorption component that absorbs the specificenergy and that is located in the coating or the core, or both; and thespecific property is the absorption of the specific energy to rupturethe microparticle, releasing the chromophore which disperses in thetissue, thereby changing or removing, or both, the detectable marking,wherein the coating, the core, or the optional absorption component, orany combination thereof, provides the specific property.

In various embodiments, chromophores can be made from any appropriatesolid, liquid, or gaseous material that has chromophoric properties. Ingeneral, useful chromophores include stains, dyes, colored drugs andproteins, and other materials. In many embodiments, chromophores arebiologically inert and/or non-toxic (ideally they are non-carcinogenic,non-allergenic, and non-immunogenic) such as those approved by the FDAfor use within the body.

In various embodiments, chromophores may be mixed in combinations beforeor after optional encapsulation, so that it may only be necessary toselect a small number of different chromophores to obtain a broad rangeof colors for various tissue marking purposes. For example, the purechromophores can be encapsulated separately and afterwards differentcolors may be mixed to form intermediate colors and shades (yellowmicroparticles may be mixed with blue microparticles to form a greenmixture). Combinations of two or more unreactive chromophores can bemixed to form desired colors and shades, and then encapsulated to formmicroparticles. Optionally, pure chromophores may be separatelyencapsulated to form sub-microparticles, and then different coloredsub-microparticles can be mixed together (or with unencapsulatedchromophores) to form desired colors and shades. The mixture can then beencapsulated in coating to form a microparticle having a perceived colorresulting from the blend of the differently colored chromophores.

In various embodiments, useful dispersible chromophores include, but arenot limited to: drugs and dyes such as rifampin (red), β-carotene(orange), tetracycline (yellow), indocyanine green (such asCardio-Green™), India ink, Evan's blue, methylene blue; solubleinorganic salts such as copper sulfate (green or blue), Cu(NH₃)²⁺ (darkblue), MnO₄ (purple), NiCl₂ (green), CrO₄ (yellow), Cr₂O₇.²⁻ (orange);proteins such as rhodopsin (purple and yellow forms) and greenfluorescent protein (fluoresces green under blue light); and any of theFood and Drug Administration (FDA) approved dyes used commonly in foods,pharmaceutical preparations, medical devices, or cosmetics, such as thewell-characterized non-toxic sodium salts FD&C Blue No. 1 (BrilliantBlue FCF), FD&C Green No. 3 (Fast Green FCF), FD&C Red No. 3(Erythrosine), FD&C Red No. 40 (ALLURA™ Red AC), FD&C Yellow No. 5(Tartrazine), and FD&C Yellow No. 6 (Sunset Yellow FCF). Of these FD&Cdyes, Yellow No. 5 is known to produce occasional allergic reactions.Additional FDA approved dyes and colored drugs are described in the Codeof Federal Regulations (CFR) for Food and Drugs (see Title 21 of CFRchapter 1, parts 1-99).

In various embodiments, dispersible chromophore nanoparticles can bemade from certain inert, normally indispersible colored substances thathave been reduced to nanoparticles about 50 nm and smaller (e.g., 0.1-5nm, 5-25 nm, 25-50 nm, and overlapping ranges therein). Although diffusenanoparticles might have different optical properties from themacroscopic material, when concentrated within the confined space of amicroparticle core (that is, nanoparticles are closer together than thewavelength of visible light, about 500 nm), they act as a single lightscatterer and/or absorber, and thus have the appearance of the originalindispersible material from which they are derived. Useful dispersiblechromophore nanoparticles may be made from graphite, iron oxides, andother materials with small particle size, for example, less than 50 nm,less than 5 nm, etc.

In some embodiments, like the coating material, chromophores can be amaterial, or can include specific absorption components, which stronglyabsorbs radiation of specific wavelength(s), particularly in thenear-infrared spectral region from about 800 to 1800 nm. Absorptionproperties of the chromophore or specific absorption component allow themicroparticle core to be selectively heated by pulses of near-infraredradiation, thus rupturing the microparticle and releasing the previouslyencapsulated chromophores.

Visibly colored near-infrared absorbing materials can be used as thechromophore(s) (to provide the desired detectable color) or as specificabsorption component(s) in conjunction with another chromophore (tocontribute to the detectable color, if desired). The infrared-absorbingvisible chromophore should be rendered invisible upon exposure of themicroparticles to the radiation, for example, through dispersal.Examples of useful colored near-infrared absorbing materials include,but are not limited to, graphite and amorphous forms of carbon (black),iron oxides (black or red), silicon (black), germanium (dark gray),cyanine dyes (including indocyanine green and other colors),phthalocyanine dyes (green-blue), and pyrylium dyes (multiple colors).See also U.S. Pat. No. 5,409,797, herein incorporated by reference.

Near-infrared absorbing materials used as specific absorptioncomponent(s) can also be visibly transparent or nearly transparent atthe concentrations and sizes used within the microparticles so that theydo not affect the perceived color of the microparticle or of the tissueafter microparticle disruption even if the material is indispersible.Useful examples include particles of filter glass (such as thosemanufactured by Schott, Inc.) and plastics such aspolymethylmethacrylate (PMMA), as well as low concentrations ofnanoparticulate graphite or other carbon. These materials can be mixedwith chromophores having a desired color and then encapsulated.

In various embodiments, materials with other properties (such asabsorption of ultraviolet, visible, microwave, radio wave and otherwavelengths) can also be used to construct the photoactive materials.For example, visible materials can be incorporated into themicroparticles as chromophores, or as specific absorption componentswithin the chromophore or coating material. Then visible radiation canbe applied to rupture the microparticles. Useful materials include, butare not limited to, all of the visible colored dispersible chromophoreslisted above and other materials rendered invisible upon exposure of themicroparticles to the visible radiation, for example, Oil Nile Blue Ndyes, fluorescein dyes, porphyrin dyes, and coumarin dyes.

In another embodiment, chromophores can be materials that are renderedinvisible (or whose color changes) upon exposure of the microparticlesto specific electromagnetic radiation without necessarily rupturing themicroparticle. Bleachable chromophores (which react with a bleachingagent released by the radiation), photobleachable chromophores (alteredby the radiation) or thermolabile chromophores (altered by heat producedby radiation absorption) may be used. Most of the chromophores listedabove are suitable, because they can be oxidized and rendered invisibleby bleaching agents, for example, peroxides, hypochlorites (such assodium hypochlorite, or household bleach), excited oxygen species, orfree radicals. For example, a microparticle can be constructed with corechromophore FD&C Red No. 40 and sub-microparticle(s) 90 containingsodium hypochlorite as the bleaching agent, which is released uponexposure of the microparticle to specific electromagnetic radiation. Thechromophore FD&C Red No. 40 is rendered invisible upon exposure of themicroparticle to this radiation and mixing with the bleach. Bleachablechromophores, which are pH-sensitive can also be used, because they canbe rendered invisible if the pH within the microparticle is changed. Forexample, a microparticle can be constructed with core chromophorephenolphthalein (pink to red above pH 9) in a basic alcohol solution andsub-microparticle(s) 90 containing hydrochloric acid as bleaching agent100 which is released upon exposure of the microparticle to specificelectromagnetic radiation. The chromophore phenolphthalein is renderedinvisible upon exposure of the microparticle to this radiation becauseof reduction in pH within the microparticle.

Photobleachable chromophores that are colored until they are renderedinvisible by exposure to a specific type, wavelength, and/or intensityof electromagnetic radiation include, but are not limited to,phthalocyanine (such as the zinc or chloroaluminum complexes which aregreen or blue); porphycenes which can be green or purple; chlorin whichis a chlorophyll derivative; rhodamine dyes which can appear red,yellow, or orange and are bleached upon exposure to near-ultravioletlight; porphyrins (such as porfimer sodium, for example, PHOTOFRIN™(Quadra Logic Technologies, Vancouver, British Columbia, Canada), agreen chromophore bleached by near-ultraviolet light); Rose Bengal,bleached upon exposure to near-ultraviolet light or high intensityvisible light (such as in the megawatts/cm² range); andinfrared-bleached dye-paired ion compounds, cationic dye-borate anioncomplexes, 3-position-substituted coumarin compounds, andbis(diiminosuccinonitrilo)-metal complexes, as described in U.S. Pat.No. 5,409,797, herein incorporated by reference. Some chromophores areonly photobleached upon simultaneous absorption of multiple photons, andare therefore unaffected by diffuse solar radiation.

Formulations. In several embodiments, photoactive materials may belipophilic or non-lipophilic. Generally, a lipophilic photoactivematerial is dissolved in a pharmaceutically acceptable oil and applieddirectly to the area of skin one wishes to treat. A lipophilicphotoactive material is dissolved in oil at a final concentration asdescribed herein. As provided herein, the photoactive materials areformulated into a non-dispersive composition in order to retain thephotoactive materials at the target region. In certain embodiments, thenon-dispersive composition contains at least one of water, a humectant,a surfactant, a thickener, a dye, an antiseptic, an anti-inflammatoryagent, an anti-oxidant, a vitamin, a fragrance, an oil, or a topicalanesthetic.

In other embodiments, the thermal damage to the epidermis resulting fromthe photoactive material reduces the efficacy of the barrier function ofthe epidermis, in particular decreasing the stratum corneum. Thisfacilitates the delivery of drugs or other substances to the dermis andepidermis, which can either enhance the effects of the treatment, ordecrease the side effects caused by partial damage of the epidermisand/or dermis, or both. Such beneficial ingredients (such as drugs andother substances), which may enhance the efficacy of skin remodelinginclude, but are not limited to, growth factors, collagen byproducts,collagen precursors, hyaluronic acid, vitamins, antioxidants, aminoacids, retinoids, retinoid-like compounds, and supplemental mineralsamong others. Groups of drugs and substances, which may decrease sideeffects, can be steroidal anti-inflammatory drugs, non-steroidalanti-inflammatory drugs, antioxidants, antibiotics, antiviral drugs,antiyeast drugs and antifungal drugs. In an embodiment of the presentinvention, the vitamins that are used may be vitamin C and/or vitamin E.The supplemental minerals used may be copper and zinc. The antioxidantscan be, for example, vitamin C and/or vitamin E. Skin lightening,whitening, or brightening agents are also provided. In severalembodiments, one or more of the ingredients described herein areincluded in the same formulation as the photoactive particles. In otherembodiments, these ingredients are provided after treatment with thephotoactive particles. In one embodiment, the efficacy of theseingredients are enhanced when used in combination with the photoactiveparticles.

In further embodiments the epidermis may be treated with sealing orbonding agents to restore barrier function on the skin to preventinfection or scarring. Alternatively bonding agents can be used totighten, pull, bond, close or otherwise change the mechanical forceswithin or around the lesion or scar (e.g. reducing tension) during orafter treatment to direct the wound healing response, including thedeposition of new collagen and re-epithelialization in response totension. Sealing or bonding agents known in the art include, but are notlimited to, cyanoacrylates and other adhesives.

In order to provide effective dermal penetration into the target tissue,the photoactive particles (e.g., plasmonic nanoparticles) in certainembodiments are formulated in various compositions. Preferentially, thenanoparticles are formulated in compositions containing 1-10% v/vsurfactants (e.g. sodium dodecyl sulfate, sodium laureth 2-sulfate,ammonium lauryl sulfate, sodium octech-1/deceth-1 sulfate). Surfactantsdisrupt and emulsify sebum or other hydrophobic fluids to enableimproved targeting of hydrophilic nanoparticles to a target region ofthe skin containing a scar or other feature to be targeted fortreatment. Surfactants also lower the free energy necessary to deliverhydrophilic nanoparticles into small hydrophobic crevices. Compositionsmay also include emulsions of particles at various concentrations (1-20%w/v) in aqueous solutions, silicone/oil solvents, propylene glycol orcreams (e.g. comprising alcohols, oils, paraffins, colloidal silicas).In other embodiments, the formulation contains a degradable ornon-degradable polymer, e.g., synthetic polylactide/co-glycolideco-polymer, porous lauryllactame/caprolactame nylon co-polymer,hydroxyethylcellulose, polyelectrolyte monolayers, or alternatively, innatural hydrogels such as hyaluronic acid, gelatin and others. Infurther embodiments, a hydrogel PLGA, PEG-acrylate is included in theformulation. Alternatively, a matrix component such as silica,polystyrene or polyethylene glycol is provided in the formulation. Otherformulations include components of surfactants, a lipid bilayer, aliposome, or a microsome. A particle may comprise a larger micron-sizedparticle.

Applicator devices. A benefit of various embodiments of the presentinvention is the effective treatment of scar tissue, while minimizingadjacent epidermal tissue damage. In various embodiments, thephotoactive materials are formulated for dispensing (including but notlimited to from an applicator device 300) in a volume of less than about1 nanoliter to greater than 100 microliters, such as 1 nanoliters, or10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160,170, 180, 190, 200, 210, 220, 230, 240, 250, 300, 350, 400, 450, 500 orabove about 500 nanoliters, such as 600, 700, 800, 900 nanoliters, or 1,2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21,22, 23, 24, 25, 30, 35, 40, 45, 50 or above about 50 microliters, suchas 75, 100, 150, 200, 250, 300, 400, 500, 600, 700, 800, 900, 1000, orabove 1000 microliters, and any ranges between any numbers therein.

In one embodiment, such formulations are in viscous compositions (hereintermed “non-dispersive compositions”) such that lateral movement acrossthe surface of the skin is reduced or prevented. A non-dispersivecomposition is substantially retained at the epidermal site ofapplication for a period of time sufficient for the light source to bedirected to the target region and one or more exposures of the lightsource to be completed.

In one embodiment, the apparatus is capable of delivering a volume of anon-dispersive composition of from about 100 nanoliters to about 50microliters of the liquid formulation on the target area such that thesurface area of the target area contacted by the liquid formulation isless than about 50 mm², e.g., 25 mm² or less.

In one embodiment, the non-dispersive composition in a volume of about0.1-2 microliters (e.g., about 0.5 microliters) has a diameter of lessthan about 2-10 m, (e.g., about 5 mm) at the epidermal surface for atleast one minute after contacting of the non-dispersive composition withthe epidermal surface. In several embodiments, the photoactive materialdoes not substantially penetrate the epidermal surface.

In one embodiment, the non-dispersive composition does not laterallymigrate along the epidermal surface upon which it has been applied at arate greater than about 1 mm per minute.

In one embodiment, a thickener is added to the composition to achieve aviscosity that provides non-dispersive properties to the formulation.

In some embodiments, the formulations are dispensed using a needle-noseor fine tip applicator, such as those used for dispensing epoxy or otherviscous materials. Alternatively, the formulations are dispensed using apipette, such as a glass or plastic-tipped pipet.

In certain embodiments, an apparatus for delivering a formulation intoan acne scar of a human subject contains a supply of a liquid (orsemi-liquid such as a gel, or semi-solid such as a paste) formulationcontaining an photoactive material, which, when put in substantialphysical contact with a target area of a skin surface of a humansubject, is capable of penetrating the skin surface at the target area(such as an acne scar) to denature at least one pathophysiologicalcollagen deposition present in the acne scar. The supply of theformulation containing the photoactive material may contain sufficientmaterial for a single administration or multiple administrations (e.g.,to a single target region multiple times or to multiple target regionsone or more times).

Other embodiments for the targeted delivery of photoactive material tosmall target regions of dermal scar tissue will be apparent to thoseskilled in the art, in particular those versed in the field of make-upartistry. Applicators for the delivery of photoactive material myinclude, but are not limited to, a pen, pencil, stencil, brush, powder,metered dosing applicator, sponge, cloth, hand, spray device and otherapplicators known in the field.

Further provided herein are systems containing an apparatus and a lightsource. In several embodiments, systems are useful for treating acnescars by denaturing collagen present in an acne scar of a human subject,containing a light source (e.g., a laser light) and an apparatus thatcontains a supply of a liquid formulation containing a photoactivematerial, which, when a volume of from about 0.01 ml to about 1 ml isapplied in substantial physical contact with a target area of a skinsurface of a human subject, is retained in the target area and iscapable of penetrating the skin surface at the target area to denatureat least one pathophysiological collagen deposition present in the acnescar, by delivering sufficient thermal energy to the targeted area suchthat the temperature of the collagen deposition in the target area iselevated above the denaturation temperature of the collagen deposition.

In some embodiments the formulations are applied to larger areas beyondthe target area, either by painting, swabbing, spraying, or pouring thephotoactive formulations on this larger area, and then the formulationis removed, such as by wiping, blotting, suction, or otherwise, from thenon-target regions. In some embodiments, an adhesive material is appliedto the target skin region in order to enhance retention of thephotoactive material at the target skin. Alternatively, a material isapplied to a non-target region that diminishes or prevents thephotoactive material from adhering to the area outside of the targetregion.

In some embodiments compositions are formulated to enable photoactivecompounds to be easily removed from non-target regions or domains afterinitial application. For example, “non-sticky” surface coatings may beapplied to photoactive material to reduce binding to the skin surfaceand/or components thereof. Coatings include, but not limited to, silica,polyvinyl pyrrolidone, polysulfone, polyacrylamide, polyethylene glycol,polystyrene cellulose, carbopol or other polymers, monolayers, orcompounds that modify charge, alter hydrophobicity, render a surfacealiphatic, or otherwise change the binding nature of a material with theskin. In other embodiments the carrier solution may be modified by theaddition of surfactants or solvents that change the binding propertiesof photoactive materials within the formulation.

In some embodiments a device is used to redistribute photoactivematerial once applied. This redistribution may include expanding thecoverage area of the applied composition, increasing the depth ofpenetration of material in a small crevice or pitted scar, removingvolume from applied composition, removing material from non-target skinareas or other patterns of redistribution. Devices that can beadvantageous for redistributing material include, but are not limitedto, a swab, brush, sponge, cloth, wipe, knife, fine tip applicator andother devices known in the art.

In one embodiment, the mixture is contacted with the skin for about 1minute to about 1 hour prior to irradiation, though embodiments whereinthe solution is contacted beyond 1 hour are provided. In one embodiment,the radiation is administered using a laser capable of delivering one ormore wavelengths.

While the invention can be performed, according to several embodiments,using intense pulsed light (IPL) systems, in general narrower range(s)of wavelengths are administered by the laser as compared to broadwavelength ranges delivered with intense pulsed light devices. Theadministration of more discrete wavelengths permits more accuratecontrol of laser effects than is easily performed when using broadspectrum IPL sources, and moreover, the accurate determination of theproper amount of energy to be provided to a target skin region of apatient is easier with a laser than with and IPL source.

In one embodiment, the energy can be tuned by monitoring thermal heatgradients on the surface of the skin with a thermal/infrared camera. Asdemonstrated herein, the methods and systems of the present disclosureprovide superior efficacy when a surface plasmon is generated on thenanoparticles by the action of the radiation. Typically, the plasmon isgenerated in a one-photon mode or, alternatively, a two-photon mode, amulti-photon mode, a step-wise mode, or an up-conversion mode.

In various embodiments, the target region is exposed to light of afrequency, for a duration, and for a number of repetitions to provide anamount of heating of all or a substantial portion of the target regionthat is sufficient to heat at least a portion of the dermal scar tissueto a temperature of at least 40 degrees Celsius, such as 45, 50, 55, 60,65, 70, 75, 80 or above 80 degrees Celsius for a period of timesufficient to be effective, meaning to cause lethal damage and/orsublethal damage to surrounding parts of the target region, and toreduce the dermal scar tissue. Provided herein are single or multipleexposures useful to achieve the appropriate thermal damage in particulartarget regions.

The optical source may be coupled to a skin surface cooling device toreduce heating of particles or structures on the skin surface outside ofthe target region to thereby focus heating to a target region. Forexample, the treatment incorporates some form of epidermal cooling,which can be administered to the entire face or entire cosmetic unit(e.g., the cheek, chin, nose, or forehead). In various embodiments,cooling devices may include, but are not limited too, refrigerated air,forced air, cryogen spray, cryogen based dynamic cooling, contactcooling (e.g. sapphire window), and other skin surface cooling systemsknown in the art.

Methods of treatment. In some embodiments, the photoactive materials areapplied non-dispersively to a target region of skin that has anepidermal surface and dermal scar tissue, which typically contains asingle acne scar. In one embodiment, the target region does not exceedabout 25 mm². Energy in the 700 nm to about 1200 nm range is deliveredto the target region in an amount sufficient to heat at least a portionof the dermal scar tissue to a temperature of at least 40 degreesCelsius for a period of time sufficient to reduce the dermal scartissue.

In another embodiment, provided are methods for reducing dermal scartissue in a human subject, that involve first identifying a targetregion of skin tissue on a human subject, where the target regioncomprises an epidermal surface and dermal scar tissue comprising apathophysiological collagen deposition, dermal matrix, or epidermalsurface, and generally where the target region does not exceed about 25mm². Pre-identification and selection of the target region is asignificant advantage to the present invention as it prevents orsubstantially reduces injury to non-target regions, thus increasingefficacy, patient comfort, and healing time. The epidermal surface ofthe identified target region is contacted with a non-dispersivecomposition containing a photoactive material, and energy is deliveredto the target region in the 700 nm to about 1200 nm range in an amountsufficient to heat at least a portion of the dermal scar tissue to atemperature sufficient to cause damage and regeneration, therebyreducing the dermal scar tissue.

In some embodiments, provided are methods for preventing the formationof dermal scar tissue or reducing its progression that involveidentifying target regions of inflammatory acne lesions on a humansubject, contacting the region with a non-dispersive compositioncontaining photoactive material, and delivering energy to the targetregion in the 700 nm to about 1200 nm range in an amount sufficient toheat at least a portion of the inflammatory acne lesion to a temperaturesufficient to cause damage and regeneration, thereby treating theinflammatory lesion and or reducing the dermal scar tissue resultingfrom the lesion.

In several embodiments, excessive sweating (e.g., hyperhidrosis istreated) with the application of photoactive particles. In oneembodiment, the method includes i) topically administering to a skinsurface of the subject a composition of photoactive particles (e.g.,plasmonic particles) ii) providing penetration means to redistribute theplasmonic particles from the skin surface to the sweat glands (e.g.eccrine sweat glands, apocrine sweat glands) and iii) causingirradiation of the skin surface by light to activate photoactivematerials and thereby heat, damage, treat or otherwise modulate thesweat gland to reduce excessive sweating.

The application of the photoactive material and delivery of energythereto may be performed once or may repeated one or more times on thesame target region, or alternatively, to one or more additional targetregions. While the target region of skin can be located anywhere on thehuman subject's body, acne vulgaris scars are most prominent on the faceor neck of the human subject.

EXAMPLES Example 1. Generation of Plasmonic Nanoparticles forThermomodulation

In one embodiment, plasmonic nanoparticles, including nanorods, hollownanoshells, silicon nanoshells, nanoplates, nanorice, nanowires,nanopyramids, nanoprisms, nanoplates and other configurations describedherein and known to those skilled in the art, are generated in sizeranges from 1-1000 nm under conditions such that surface properties thatfacilitate deep follicular penetration. Surface properties can be variedon one or multiple (2, 3, or 4) different dimensions to increasenanoparticle concentration in a target tissue domain. Penetration intofollicular openings of 10-200 um can be maximized using thenanoparticles described herein. Here, nanoparticles sized in the rangeof about 10 to about 100 nm are generated, and are preferably assembledor formulated into multiparticle structures having a size in the rangeof 100-300 nm. Alternatively, a coating (e.g., silica) is grown onuniparticular structures to increase the particle size to the range of100-300 nm or more.

Surface-modified plasmonic nanoparticles. An embodiment of a preparationof surface-modified plasmonic nanoparticles is provided as follows.Plasmonic nanoparticles are synthesized with stablecetryltrimethylamonium bromide (CTAB) coating and concentrated from anoptical density of 1 O.D. to 100, 200, 300, 400, or 500 O.D. through oneto three cycles of centrifugation at 16,000 rcf, with supernatantdecanting. Alternatively, CTAB-coated nanoparticles are concentrated andresuspended in 250 Amol/L 5-kDa methyl-polyethylene glycol (PEG)-thiolto make PEG-coated nanoparticles. Verification that PEG polymer stocksare fully reduced is performed using spectrophotometry to measure thethiol activity of polymer-thiols with 5,5-dithiobis(2-nitrobenzoic acid)against a DTT gradient. The solution of methy-PEG-thiol and CTAB-coatednanoparticles is mixed at room temperature for 1 h then dialyzed against5 kDa MWCO in 4 L distilled water for 24 h. Dialyzed samples areprocessed through 100-kDa filters to remove excess polymer.Quantification of the number of PEG polymers per particle is performedby surface-modifying nanoparticles with amino-PEG-thiol polymer andquantifying the number of amines with an SPDP assay. For testformulations, 100 O.D. solutions of CTAB-coated plasmonic nanoparticlesare made in distilled water, and 100 O.D. PEG-coated plasmonicnanoparticles are made in distilled water, ethanol, DMSO, or mineraloil. Plasmonic nanoparticles with silica shells are created by reactingnanoparticles with silicates such as tetra-ethyl-ortho-silicate (TEOS),sodium silicate, aminopropyletriethoxysilane (APTS), etc. to thicknessesof 5-50 nm or more. Control, vehicle-only formulations contain nonanoparticles.

Embedded nanoparticles. In one embodiment, nanoparticles are embedded(or encapsulated) in materials, which allows for the generation of adiverse range of sizes to tune their size. Particle sizes in the rangeof 100-2000 nm or 200-2000 nm have been shown to enter the hair folliclewithout penetrating the dermis. Nanoparticles are encapsulated insilica, a synthetic polylactide/co-glycolide co-polymer, porouslauryllactame/caprolactam nylon co-polymer, hydroxyethylcellulose,polyelectrolyte monolayers, or alternatively, in natural hydrogels suchas hyaluronic acid, without significantly altering plasmon resonanceproperties. Nanoparticles are embedded within 100-2000 nm materials or200-2000 nm materials without covalent attachment or by cross-linking ofamines, carboxyls or other moieties on the nanoparticle surface to thepolymer structure. The surface of the 100-2000 nm material or 200-2000nm material may be modified for an optimal zeta potential,hydrophilicity/hydrophobicity, and/or adsorption layer throughtechniques described herein. Furthermore, the shape of the aspect ratioof the polymer can be modified from low to high to increaseconcentrations and depths of penetration of the embedded plasmonicnanoparticles. The nanoparticles advantageously have an aspect ratiogreater than about 1.

Example 2. Formulation of Thermoablative Plasmonic Nanoparticles forTopical Delivery

In another embodiment, nanoparticles are generated as in Example 1 usingan appropriate solvent (e.g., water, ethanol, dimethyl sulfoxide). Themixture comprising a plurality of nanoparticles in water is concentratedto about 100-500 O.D. and exchanged for a new solvent by liquidchromatography, a solvent exchange system, a centrifuge, precipitation,or dialysis. The solvent may include an alcohol (e.g., n-Butanol,isopropanol, n-Propanol, Ethanol, Methanol), a hydrocarbon (e.g.,pentane, cyclopentane, hexane, cyclohexane, benzene, toluene,1,4-Dioxane), chloroform, Diethyl-ether, water, an acid (e.g., aceticacid, formic acid), a base, acetone, dimethyl sulfoxide (DMSO),dimethylformamide (DMF), acetonitrile (MeCN), tetrahydrofuran (THF),dichloromethane (DCM) or ethylacetate. The new solvent is combined witha cosmetically or pharmaceutically acceptable carrier, thereby forming ananoparticle composition. Generally, the particles and carrier will forman emulsion.

Plasmonic nanoparticle formulations are provided that amplify orexpedite the penetration of nanoparticles into hair follicles. In someembodiments, nano- and micro-emulsions facilitate partitioning withinlipid-rich skin compartments such as the hair follicle. In someembodiments, nanoparticles are formulated in compositions containing0.5-2% v/v surfactants to enable disruption of the epidermal skinbarrier, emulsification of sebum, and improved mixing of hydrophilicnanoparticles in hydrophobic solutions or targeting to hydrophobic spacein the skin (e.g. between the hair shaft and surrounding follicle).Formulations of nanoparticles are also provided at variousconcentrations (1-20% w/v) in aqueous solutions, silicone/oil solvents,polypropylene gel, propylene glycol or creams (e.g. containing alcohols,oils, paraffins, colloidal silicas). In some embodiments,light-absorbing nanoparticles are utilized in solutions having tailoredpH, temperature, osmolyte concentration, viscosity, volatility, andother characteristics to improve light-absorbing nanoparticle entry intohair follicles.

Formulations are prepared to maximize nanoparticle stability (degree ofaggregation in solution), nanoparticle concentration, and nanoparticleabsorbance (degree of laser-induced heating at differentconcentrations).

When formulations of plasmonic nanoparticles are illuminated with aclinical laser with a wavelength coincident to the peak absorptionwavelength of the particle, the formulation heats to thermoablativetemperatures more rapidly and to a greater degree than conventionalclinical absorptive dyes. FIG. 2 compares the temperature profile ofplasmonic particles (1020 nm peak absorption wavelength) to conventionalclinical dyes carbon lotion, meladine spray and indocyanine green afterexposure to 1064 nm, 20 J/cm², 55 ms laser pulses. The temperatureincrease caused by pulsed 1064 nm laser light was more than 2.5 timesgreater for the plasmonic solution, compared to conventional clinicaldyes used at the same dilution (1:1000 dilution from clinicalconcentration, where clinical concentrations are as follows: carbon20-200 mg/ml, meladine 1 mg/ml, indocyanine green 5 mg/ml).

Example 3. Use of Plasmonic Nanoparticles for Thermomodulation of Hair

Individuals having blonde, red, gray, or lightly-colored hair are notadequately treated with existing traditional light-based hair removaltechniques. Provided herein are methods for using the compositionsdescribed herein for the selective removal or reduction of untreatedblonde, red, gray, or lightly-colored hair. In one embodiment, plasmonicnanoparticles generated and formulated as described above are introducedinto a target tissue region, generally a skin region, and activated withlaser-based hair removal systems as known in the art in order to achieveeffective hair removal.

To achieve maximal penetration depth and concentration of plasmonicnanoparticles in the hair follicle and/or near components of thesebaceous gland including the sebaceous duct, the sebum, the epitheliallinking of the sebaceous gland, and/or near the bulge region includingthe stem cells, stem cell niche, epithelial lining of the bulge region,and/or near the follicular bulb, an optimal particle size of 30-800 nm(e.g., 100-800 nm) containing one or several plasmonic nanoparticles isconstructed. Nanoparticles encapsulating plasmonic nanoparticles can beformulated from any number of polymers or matrices. In some embodiments,the formulation contains a degradable or non-degradable polymer, e.g.,synthetic polylactide/co-glycolide co-polymer, porouslauryllactame/caprolactame nylon co-polymer, hydroxyethylcellulose,polyelectrolyte monolayers, or alternatively, in natural hydrogels suchas hyaluronic acid, gelatin and others. In further embodiments, ahydrogel PLGA, PEG-acrylate is included in the formulation.Preferentially, a matrix component such as silica, polystyrene orpolyethylene glycol is provided in the formulation to improve particlestability and enable facile removal from the skin surface afterapplication and follicle targeting. Other formulations include componentof surfactants (e.g. sodium dodecyl sulfate, sodium laureth 2-sulfate,ammonium lauryl sulfate, sodium octech-1/deceth-1 sulfate), a lipidbilayer, a liposome, or a microsome. Plasmonic nanoparticles includingnanorods, nanoshells, nanospheres, nanoplates, or nanorice can beencapsulated within a the polymer or lipid-based nanoparticle or matrixor deposited on the particle surface. Alternatively, nanoparticles inthe size range of 100-250 nm, 250-500 nm, 800 nm-1500 nm, or greaterthan 1500 nm can be used.

Pre-treatment of skin with mechanical or chemical exfoliation is used insome embodiments to remove hair-plugs and “open” the follicle forparticle delivery. Additionally, hairs can be shaven or waxed to createa void in the hair follicle for particles to fill. The use of physicalor thermal force amplifies or expedites the penetration of lightabsorbing nanoparticles and conjugates thereof into hair follicles, inpart by causing dilation of the hair follicle prior to application ofthe nanoparticles. For example, ultrasound and other sonic forces,mechanical vibrations, hair shaft manipulation (including pulling),physical force, thermal manipulation, and other treatments are utilizedto improve entry of light-absorbing nanoparticles into hair follicles.Nanoparticle formulation treatments are performed alone, in combination,sequentially or repeated 1-24 times.

An applicator is used to uniformly apply the composition ofnanoparticles into follicles. The applicator can be a sponge, a cloth,direct contact from a finger, a tube, a syringe, a device that appliessuction, an aerosol, a spray, or other means known in the art. In oneexample, a formulation of 1 ml of plasmonic nanoparticles at aconcentration of 100 O O.D. with peak resonance of 810 nm is applied toapproximately 200 cm² area of the skin of an adult human subject with asyringe. A cloth is used to evenly distribute solution across the skinarea and into the hair follicles. Deep massage from a mechanicalvibrator for 2 minutes with or without 1 MHz ultrasound for 5 minutes,is applied to drive particles deep into the follicle. Particlespenetrate 50-75% down the full length of the hair shaft atconcentrations sufficient to heat skin in a 100 μm radius at incrementaltemperatures of 5-20-fold greater than is generated in similar volumesof adjacent skin when irradiated by a Diode (810 nm) laser. Acetone,ethanol, or a debriding agent can be used to remove all particles fromthe surface of the skin that have not deposited in the follicle, inorder to reduced or prevent non-follicular heating of the skin.

Nanoparticle formulations are tested in ex vivo animal samples, ex vivohuman skin samples, and in vivo human skin including the assessmentof: 1) depth of nanoparticle penetration into hair follicles; 2)particle concentration achieved; 3) degree of heating achieved atdelivered nanoparticle concentrations; and 4) efficacy of photothermaldestruction including temporary and permanent hair removal, 5) clearanceof nanoparticles after treatment. To assess nanoparticle penetrationdepths, plasmonic nanoparticles surface-functionalized with fluorescentmolecules are visualized by fluorescence microscopy after histologicalsectioning or follicular biopsy (removal of hair shaft). Alternatively,plasmonic nanoparticles are directly visualized by dark field microscopyafter histological sectioning or follicular biopsy. To assessnanoparticle concentrations at various depths along the follicle,excised skin samples are separated by tape stripping or heat-basedtechniques, samples are dissolved for bulk analysis of metalconcentration by ICP-MS (inductively coupled plasma-mass spectrometry).The macroscopic degree of heating is validated by infrared thermographyof skin samples, and by assessment of skin sections subject to laserexposure for thermal damage markers. Finally, one can measure efficacyof photothermal destruction at the nanoparticle accumulation site byanalyzing histological cellular lesions at the target site, includingthe follicular hair shaft, inner root sheath, outer room sheath, andbulge region containing the stem cell niche, which contains the stemcells that contribute to new hair growth. As the bulge region isgenerally localized about midway (˜50% down the length of) the hairshaft, permanent hair removal is sufficiently achieved by accumulationof plasmonic nanoparticles to this depth. In some situations,nanoparticle delivery may also generate a heat gradient emitting furtherdown the hair shaft. Animal studies are useful to demonstrate theefficacy of unpigmented hair removal by comparing heat profiles, thermalablation of hair shaft, and thermal damage of bulge stem cells intreated hairless rodents, albino rodents and dark-haired rodents.Efficacy on live human skin is measured by measuring hair counts at 3and 12 month follow ups. Biopsies are taken from select patients at 2,4, and 6 week follow ups to verify that nanoparticles are cleared fromthe skin without embedding in the dermis.

Hair follicle penetration of fluorescently-labeled nanoparticlesdetermined using porcine skin explants and confocal imaging. A 25 mg/mlaqueous solution silicon dioxide-coated nanoparticles (200 nm diameter)was contacted with freshly thawed porcine skin, after which excessnanoparticle suspension was removed and manual massage was performed forthree minutes. The explant was sectioned and subjected to confocalimaging. As shown in FIG. 3A, explant sections were imaged at angles tothe hair follicles in 60 μm planes; Plane 1 shows the follicleinfundibulum, while Plane 2 shows the distal regions of the follicle.FIG. 3B demonstrates representative confocal images showing that rednanoparticles (548 nm absorbance) are visible within both thesuperficial and deep follicles, but are not detectable in dermal layersbeneath the follicles. FIG. 3C shows high-magnification imaging of rednanoparticles localized to and retained within a deep follicle (˜400μm). Green color indicates tissue autofluorescence (488 nm).

Hair follicle penetration of plasmonic nanoparticles determined usingporcine skin and dark field imaging. A 100 O.D. suspension of plasmonicnanoparticles (200 nm diameter) was contacted with freshly thawedporcine skin, after which excess nanoparticle suspension was removed andmanual massage performed for three minutes. The procedure was repeatedfor a total of 3 applications, and surface residue removed with several3-5 applications of alternating water and ethanol. The skin sample wasexcised, fixed, sectioned along horizontal plane and subjected to darkfield imaging. As shown in FIG. 4A, skin samples were sectioned andimaged horizontal to the hair follicle at various depths. In skinsection images, plasmonic nanoparticles were observed as bright bluecolor point sources at depths up to 1.2 mm deep in porcine folliclespaces (FIG. 4B). Control samples with no plasmonic nanoparticles wereclearly differentiated (FIG. 4C). ICP-MS is also performed on skinsections to assess nanoparticle concentrations at various depths alongthe follicle.

Hair follicle penetration of nanoparticles in hairless rodents, albinorodents and dark-haired rodents. White-haired Swiss Webster mice (n=3)at 8 weeks old are anesthetized with injectable ketamine/xylazineanesthetic solution and dorsal back skin and hair washed and dried.Prior to formulation administration, three 10 cm×10 cm areas aredemarcated by permanent marker on each mouse and subjected to hairremoval by 1) electric razor, 2) Nair depilation reagent, or 3) warmwax/rosin mixture application and stripping. Each mouse is treated bypipette with up to 3 nanoparticle formulations, in quadruplicate 5-μlspot sizes per demarcated skin area (up to 12 spots per area or 36 spotsper mouse). Precise spot locations are demarcated with pen prior topipetting. Duplicate treatment spots on the dorsal left side aremassaged into skin for 5 minutes, while duplicate treatment spots on thedorsal right side are applied without massage. Thirty minutes afterapplication, mice are sacrificed by carbon dioxide asphyxiation andcervical dislocation, and skin is carefully excised and punched intosections along spot size demarcations. Skin biopsies are fixed in 10%paraformaldehyde, paraffin-embedded, and cut into 5-um sections on amicrotome in transverse directions. Slides with mounted paraffinsections are deparaffinized and stained with hematoxylin and eosin (H&E)or kept unstained for dark field microscopy. Using H&E staining, lightmicroscopy and/or dark field microscopy, greater than 50 follicles performulation are imaged, and scoring is performed for skin sections forvisible macroscopic nanoparticle accumulation in the follicle, along thehair shaft, at the site of the putative bulge stem cell niche, and atthe depth of the follicle bulb. On serial histological sections, asilver enhancement staining kit based on sodium thiosulfate may be usedto enlarge the plasmonic nanoparticle signal via the precipitation ofmetallic silver. Phase and dark field micrographs are captured and usedto record the depths of follicular penetration for each nanoparticleformulation and method of application. ICP-MS is also performed on skinsections to assess nanoparticle concentrations at various depths alongthe follicle.

Assessment of photothermal destruction at the nanoparticle accumulationsite. Treated areas of pig, human or mouse skin are irradiated with alaser coincident with the peak absorption wavelength of nanoparticles(e.g. 1064 nm YAG laser for 1020 nm plasmonic particles) using clinicalparameters (1 s exposure of 30-50 J/cm² and a pulse width of 10-50 ms).To determine microscopic photothermal damage of target skin structuressuch as the hair follicle and hair follicle bulge stem cells, at tendays after application and irradiation, human subjects receive lidocaineinjections to numb treatment areas and skin is carefully excised andpunched into sections along spot size demarcations. Fresh human skinbiopsies or explanted human and animal skin samples are fixed in 10%paraformaldehyde, paraffin-embedded, and cut into 5-um sections on amicrotome in transverse directions, or they are fixed in Zamboni'ssolution with 2% picric acid and cryosectioned by freezing slidingmicrotome. Slides with mounted paraffin sections are deparaffinized andstained with hematoxylin and eosin (H&E). Histological sections areexamined at various depths for markers of thermal damage andinflammation. Hematoxylin and eosin (H&E) is used to image skin andfollicle microanatomy and indicate degeneration of hair shafts, atrophyof sebaceous glands, and cell vacuolization (indicating cellulardamage). Nitro blue tetrazolium chloride (NBTC), a lactate dehydrogenasestain that is lost upon thermal injury to cells, is used to assessdamage to keratinocytes. Cellular damage in follicles of skin samplesreceiving plasmonic nanoparticle plus laser treatment is scored andcompared to those receiving laser treatment alone. Live treated humanskin areas are also followed clinically for 2 weeks to 3 monthsfollowing plasmonic nanoparticle+laser treatment, or during repeatedplasmonic nanoparticle+laser treatments, and compared to baselinedigital photograph taken prior to first treatment, and to negativecontrol laser only treatments. Clinical observations of hair removal, aswell as erythema, edema, discomfort, irritation or scarring, are notedto determine degree of non-specific thermal damage.

Effect of plasmonic particle coating on specificity of delivery andphotothermal heating. Preferentially, a matrix component such as silica,polystyrene or polyethylene glycol is provided in the formulation toimprove particle stability and enable facile removal from the skinsurface after application and follicle targeting. Acetone, ethanol, or adebriding agent can be used to remove all particles from the surface ofthe skin that have not deposited in the follicle, in order to reduced orprevent non-follicular heating of the skin. In FIG. 5, live human skinwas treated with uncoated plasmonic particles compared to silica-coatedplasmonic particles, prior to laser-irradiation and comparison to noparticle treatment (laser only) controls. Pre-treatment of skin,including shaving with razor and microdermabrasion (15 sec, mediumsetting) to remove hair-plugs and “open” the follicle for particledelivery, was performed on both forearms. Human forearm skin wasirradiated with 810 nm laser pulses (30 J/cm², 30 ms, 2 passes) alone(FIG. 5A), or after treatment with a formulation of 830 nm resonant,Uncoated plasmonic nanoparticles in 20% propylene glycol (FIG. 5B). Theplasmonic nanoparticle formulation was applied with 3 minute massage andrepeated 3 times, and the skin surface wiped with 3 applications ofalternative water and ethanol before laser irradiation. At 30 minutesfollowing laser irradiation, non-specific clinical burns were observeddue to significant photothermal heating of residual, uncoated particleson the skin surface (FIG. 5B). Live human skin was also irradiated with1064 nm laser pulses (40 J/cm², 55 ms, 3 passes) alone (FIG. 5C), orafter treatment with a formulation of 1020 nm resonant, Silica-coatedplasmonic nanoparticles in 20% propylene glycol (FIG. 5D). The plasmonicnanoparticle formulation was applied with 3 minute massage and repeated3 times, and the skin surface wiped with 3 applications of alternativewater and ethanol before laser irradiation. At 30 minutes followinglaser irradiation, no evidence of burning of the skin or erythema wasobserved, as Silica-coated particles could be sufficiently wiped fromthe skin surface (FIG. 5D). Magnified photography of the skin areatreated with Silica-coated particles+Laser shows specific photothermaldamage (perifollicular erythema and edema) in the nanoparticle-targetedsite, without damage to surrounding or non-particle-treated tissues(FIG. 6).

Example 4. Use of Plasmonic Nanoparticles for Acne Treatment

In one embodiment, provided herein are methods for using thecompositions described herein for the treatment of acne vulgaris andother acnes and acne-like skin conditions, but the selective targetingof sebaceous follicles, particularly the sebaceous glands and/or hairfollicles. Plasmonic nanoparticles generated and formulated as describedabove are introduced into a target tissue region, generally a skinregion, and activated with laser-based systems as known in the art inorder to achieve effective hair removal.

To achieve maximal penetration depth and concentration of plasmonicnanoparticles in the hair follicle and/or near components of thesebaceous gland including the sebaceous duct, the sebum, the epitheliallinking of the sebaceous gland, and/or near the bulge region includingthe stem cells, stem cell niche, epithelial lining of the bulge region,and/or near the follicular bulb, an optimal particle size of 100-800 nmcontaining one or several plasmonic nanoparticles is constructed.Nanoparticles encapsulating plasmonic nanoparticles can be formulatedfrom any number of polymers or matrices. In some embodiments, theformulation contains a degradable or non-degradable polymer, e.g.,synthetic polylactide/co-glycolide co-polymer, porouslauryllactame/caprolactame nylon co-polymer, hydroxyethylcellulose,polyelectrolyte monolayers, or alternatively, in natural hydrogels suchas hyaluronic acid, gelatin and others. In further embodiments, ahydrogel PLGA, PEG-acrylate is included in the formulation.Preferentially, a matrix component such as silica, polystyrene orpolyethylene glycol is provided in the formulation to improve particlestability and enable facile removal from the skin surface afterapplication and follicle targeting. Preferentially, formulations includesurfactants (e.g. sodium dodecyl sulfate, sodium laureth 2-sulfate,ammonium lauryl sulfate, sodium octech-1/deceth-1 sulfate), componentsof a lipid bilayer, a liposome, or a microsome. Surfactants disrupt theepidermal skin barrier, emulsify sebum, improve mixing of hydrophilicnanoparticles with hydrophobic solutions, and reduce entropic barriersto delivering hydrophilic particles to hydrophobic regions of the skin(e.g. between the hair shaft and surrounding sheath or follicle).Plasmonic nanoparticles including nanorods, nanoshells, nanospheres, ornanorice can be encapsulated within the polymer nanoparticle or matrixor deposited on the particle surface. Alternatively, nanoparticles inthe size range of 100-250 nm, 250-500 nm, 800 nm-1500 nm, or greaterthan 1500 nm can be used.

The use of physical or thermal force amplifies or expedites thepenetration of light absorbing nanoparticles and conjugates thereof intohair follicles and/or sebaceous glands, in part by causing dilation ofthe hair follicle prior to application of the nanoparticles. Forexample, ultrasound and other sonic forces, mechanical vibrations, hairshaft manipulation (including pulling), physical force, thermalmanipulation, and other treatments are utilized to improve entry oflight-absorbing nanoparticles into hair follicles and/or sebaceousglands. Nanoparticle formulation treatments are performed alone, incombination, sequentially or repeated 1-24 times.

Prior to application of the plasmonic nanoparticles, a pre-treatmentstep of removing excess sebum from the surface of the skin may beperformed using chemical and/or mechanical means. Pre-treatment of skinwith mechanical or chemical exfoliation is used in some embodiments toremove hair-plugs and “open” the follicle for particle delivery.Additionally, hairs can be shaven or waxed to create a void in the hairfollicle for particles to fill.

An applicator is used to uniformly apply the composition ofnanoparticles into follicles. The applicator can be a sponge, a cloth,direct contact from a finger, a tube, a syringe, a device that appliessuction, an aerosol, a spray, or other means known in the art. In oneexample, a formulation of 1 ml of plasmonic nanoparticles at aconcentration of 100 O.D. with peak resonance of 810 nm is applied toapproximately 200 cm² area of the skin of an adult human subject with asyringe. A cloth is used to evenly distribute solution across the skinarea and into the hair follicles. Massage from a mechanical vibrator for2 minutes with or without ultrasound at 1 MHz for 5 minutes is appliedto drive particles deep into the follicle. Particles penetrate ˜50% downthe full length of the hair shaft at concentrations sufficient to heatskin in a 100 um radius at incremental temperatures of 5-20-fold greaterthan is generated in similar volumes of adjacent skin when irradiated bya Diode (810 nm) laser. Acetone, ethanol, or a debriding agent can beused to remove all particles from the surface of the skin that have notdeposited in the follicle, in order to reduced or prevent non-follicularheating of the skin.

Delivery of plasmonic nanoparticles to the sebaceous gland determinedusing human abdominoplasty skin and dark field imaging. The humansebaceous gland exists within the pilosebaceous unit consisting of thehair, hair follicle, arrector pili muscle and sebaceous gland. In FIG.7A, a human skin biopsy is immunostained with antibodies againstCollagen IV (basement membrane marker, blue) and PGP 9.5 (nerve marker,green) to visualize representative pilosebaceous unit microanatomy,including the hair follicle (HF), sebaceous gland (SG) and arrector pilimuscle. To deliver nanoparticles to the hair follicle and sebaceousgland, skin was first pre-treated with shaving to remove extruding hair,microdermabrasion (15 sec, medium setting) to remove hair-plugs andcorneocytes, and chemical depilation to “open” follicle microwells forparticle delivery. A 100 O.D. suspension of plasmonic nanoparticles (200nm diameter), formulated in 1% sodium dodecyl sulfate (SDS) and 20%propylene glycol (PG) was contacted with excised human abdominoplastyskin, after which excess nanoparticle suspension was removed and manualmassage performed for three minutes, followed by ultrasound (1 MHz) for5 minutes. The procedure was repeated for a total of 3 applications, andsurface residue removed with 3-5 applications of alternating water andethanol. The skin sample was excised, fixed, sectioned along horizontalplanes and subjected to dark field imaging. As assessed by dark fieldimaging of horizontal skin sections, compositions of plasmonicnanoparticles with a cosmetically acceptable carrier of 1% SDS/20% PGadministered with massage and ultrasound can be delivered 400-600 μmdeep into the human follicle and specifically into the sebaceous gland(FIG. 7B).

Cosmetic formulations for follicle and sebaceous gland delivery in humanskin. Preferentially, formulations include surfactants (e.g. sodiumdodecyl sulfate, sodium laureth 2-sulfate, ammonium lauryl sulfate,sodium octech-1/deceth-1 sulfate), components of a lipid bilayer, aliposome, or a microsome. Surfactants disrupt the epidermal skin barrierand emulsify the sebum to enable improved mixing of hydrophilicnanoparticles in hydrophobic solutions. Humectants such as propyleneglycol are used to help improve topical viscosity and maintainphysiological pH. To demonstrate the efficacy and mechanism ofembodiments of cosmetic formulations for human sebaceous gland delivery,skin was first pre-treated with shaving to remove extruding hair,microdermabrasion (15 sec, medium setting) to remove hair-plugs andcorneocytes, and chemical depilation to “open” follicle microwells forparticle delivery. Two separate 100 O.D. suspensions of plasmonicnanoparticles (200 nm diameter) were formulated in 1% sodium dodecylsulfate and 20% propylene glycol (SDS/PG) or in 1% sodiumlaureth-2-sulfate and 20% propylene glycol (SLES/PG). Formulations werecontacted with two separate excised human abdominoplasty skin samples,and massage for 3 minutes followed by ultrasound (1 MHz) for 5 min wasperformed to drive particles deep into the follicles. The procedure wasrepeated for a total of 3 applications, and surface residue removed with3-5 applications of alternating water and ethanol. The skin sample wasexcised, fixed, sectioned along horizontal planes and subjected to darkfield imaging to assess particle delivery. As assessed by dark fieldimaging of horizontal skin sections, compositions of plasmonicnanoparticles with a cosmetically acceptable carrier of 1% SLES/20%administered with massage and ultrasound can be delivered 400-600 μmdeep into the human follicle and specifically into the sebaceous gland(FIG. 8B).

Impact of massage vs. ultrasound on nanoparticle delivery to humanfollicles and sebaceous gland. In some embodiments, ultrasound and/orother sonic forces, mechanical vibrations, hair shaft manipulation(including pulling), physical force, thermal manipulation, and othertreatments are utilized to improve entry of light-absorbingnanoparticles into hair follicles and/or sebaceous glands. Mechanicalmassage improves follicular penetration through hair shaft ‘pumping’mechanisms, while ultrasound enhances transdermal drug delivery throughtemporary disruption of the skin's lipid bilayer, bubble formation, andliquid microstreaming. To characterize the effects of massage decoupledfrom ultrasound, skin was first pre-treated with shaving to removeextruding hair, microdermabrasion (15 sec, medium setting) to removehair-plugs and corneocytes, and chemical depilation to “open” folliclemicrowells for particle delivery. A 100 O.D. suspension of plasmonicnanoparticles (200 nm diameter), formulated in 1% sodium dodecyl sulfate(SDS) and 20% propylene glycol (PG), was contacted with three separateexcised human abdominoplasty skin samples. In the three treated humanskin samples, massage only was performed for 3 minutes, ultrasound only(1 MHz) was performed for 5 minutes, or massage followed by ultrasoundwas performed to drive particles deep into the follicles. In a fourthsample, no particles were applied to skin. The procedure was repeatedfor a total of 3 applications, and surface residue removed with 3-5applications of alternating water and ethanol. The skin sample wasexcised, fixed, sectioned along horizontal planes and subjected to darkfield imaging to assess particle delivery. As assessed by dark fieldimaging of horizontal skin sections, compositions of plasmonicnanoparticles with a cosmetically acceptable carrier of 1% SLES/20%administered via ultrasound deliver more plasmonic nanoparticles to theinfundibulum versus massage, albeit both mechanisms facilitate delivery(FIG. 9).

Additional plasmonic nanoparticle formulations for follicle andsebaceous gland delivery in human skin. In some embodiments, plasmonicnanoparticles include nanorods, nanoshells, nanospheres, or nanorice, orplasmonic nanoparticles encapsulated within the polymer nanoparticle ormatrix or deposited on the particle surface. Preferentially, a matrixcomponent such as silica, polystyrene or polyethylene glycol is providedin the formulation to improve particle stability and enable facileremoval from the skin surface after application and follicle targeting.To demonstrate the formulation of additional plasmonic nanoparticleshapes and concentrations for follicle, infundibulum, and sebaceousgland delivery, skin was first pre-treated with shaving to removeextruding hair, microdermabrasion (15 sec, medium setting) to removehair-plugs and corneocytes, and chemical depilation to “open” folliclemicrowells for particle delivery. Separately, 10 O.D. suspensions ofSilica-coated nanoplates, 30 O.D. suspensions of polyethylene-glycolcoated plasmonic nanorods, and fluorescent silica particles wereformulated in 1% sodium dodecyl sulfate and 20% propylene glycol.Formulations were contacted with three separate excised humanabdominoplasty skin samples, and massage for 3 minutes followed byultrasound (1 MHz) for 5 min was performed to drive particles deep intothe follicles. The procedure was repeated for a total of 3 applications,and surface residue removed with 3-5 applications of alternating waterand ethanol. The skin sample was excised, fixed, sectioned alonghorizontal planes and subjected to dark field imaging to assess particledelivery. As assessed by dark field imaging of horizontal skin sections,compositions of Polyethylene glycol (PEG)-coated nanorods (gold, 15×30nm dimension) in cosmetically acceptable carrier, administered viaultrasound and massage, were observed within the follicle infundibulumat 200 um deep (FIG. 10A). Compositions of plasmonic nanoparticles(Silica-coated nanoplates) at lower concentration (10 O.D.), wereapparent at 400-600 um deep in the follicle and in the sebaceous gland(open arrow), albeit at lower concentration than comparable particles ina similar cosmetic carrier at 100 O.D (FIG. 10B).

Assessment of photothermal destruction of sebaceous gland and targetedskin structures. Nanoparticle formulations are tested in ex vivo animalskin samples, ex vivo human skin samples, and in vivo human skin asdescribed in Example 3. One can measure efficacy of photothermaldestruction at the nanoparticle accumulation site by measuring thermaldamage to sebocytes and reduction in sebum production in the treatedsebaceous follicles. To assess photothermal destruction, human skin isfirst pre-treated with shaving to remove extruding hair,microdermabrasion (15 sec, medium setting) to remove hair-plugs andcorneocytes, and chemical depilation to “open” follicle microwells forparticle delivery. Skin is contacted with a 100 O.D. suspension of 810nm resonant plasmonic nanoparticles (200 nm diameter), and is massagedfor 3 minutes followed by ultrasound (1 MHz) for 5 min to driveparticles deep into the follicles. The procedure is repeated for a totalof 3 applications, and surface residue removed with 3-5 applications ofalternating water and ethanol. Treated human skin samples are laserirradiated with 810 nm laser (40 J/cm², 30 ms, 5 pulses), and comparedto laser only treated human skin. Human skin is biopsied, fixed inZamboni's solution with 2% picric acid, and cryosectioned by freezingsliding microtome. Slides with mounted paraffin sections aredeparaffinized and stained with hematoxylin and eosin (H&E).Histological sections are examined at various depths for markers ofthermal damage and inflammation. Hematoxylin and eosin (H&E) is used toimage skin and follicle microanatomy and indicate degeneration of hairshafts, atrophy of sebaceous glands, and cell vacuolization (indicatingcellular damage). Nitro blue tetrazolium chloride (NBTC), a lactatedehydrogenase stain that is lost upon thermal injury to cells, may alsobe used to assess damage to keratinocytes vs. sebocytes. Anintracellular stain, Oil-Red-O, may be used to determine lipid and sebumoil content in treated samples. Sebum excretion rates are measured on invivo skin at 1-3 months follow up using sebum-absorbant tapes todemonstrate functional change in sebum flow. Clearance and prevention ofacne lesions is measured by patient reported outcomes and counting acnelesions at 1-3 months follow up.

Example 5. Formulation of Thermoablative Plasmonic Nanoparticles forVascular Ablation

In one embodiment, formulations are prepared to maximize nanoparticlestability (degree of aggregation in solution), nanoparticleconcentration, and nanoparticle absorbance (degree of laser-inducedheating at different concentrations) once injected into the bloodstream. Nanoparticles are generated as in Example 1 using an appropriatesolvent. The mixture comprising a plurality of nanoparticles in water isconcentrated to about 100-500 OD at peak absorbance and exchanged for anew solvent by liquid chromatography, a solvent exchange system, acentrifuge, precipitation, or dialysis. Typical exchange solvent is 0.15mol/L NaCl, 0.1 mol/L Na phosphate buffer (pH 7.2).

Example 6. Use of Plasmonic Nanoparticles for Thermoablation ofComponent(s) of Vessels and Microvessels

In one embodiment, nanoparticle-containing compositions areadministered, typically intravascularly. Subsequent to suchadministration of plasmonic nanoparticles, a laser matched to the peakplasmonic resonance of the particles (e.g., 755 nm, 810 nm, or 1064 nm)is applied to heat nanoparticles and surrounding tissue. Pulse widths of10-100 ns, 100 ns-1 ms, 1-10 ms, 10-100 ms, 100-1000 ms or continuouswave irradiation is used to achieve thermal heat gradients and localizedheating in the vicinity of particle or particles of 20-200 nm. 200 nm-2μm, 2-20 μm, 20-200 μm, 200 μm-2 mm. Thermal gradients of 20-200 nm areachieved from individual particles. Supra millimeter thermal gradientsare achieved by the collective heat deposition of many particles inveins with diameters of several hundred microns or more. Irradiation isapplied from 1 pulse to many pulses over seconds to minutes. A coolingdevice for epidermal layers is used concomitant to irradiation to reducepain and prevent thermal damage elsewhere. Laser position, fluence,wavelength, angle of incidence, pattern of irradiation is modified toachieve irradiation of vessels at specific depths between 0-10 mm, whileavoiding heating of non-target vasculature. Alternatively, laser orlight is administered through fiber optic waveguide administered via acatheter to heat the particles in larger veins.

In one embodiment a flank of the tissue is irradiated with 2 W/cm², 810nm, 1 cm beam diameter after injection of PEG-nanorods with peak plasmonresonance at 810 nm. Thermographic imaging is used to assess surfacetemperature of tissue immediately after irradiation.

Assessment of thermal damage to component(s) of vessels, microvessels,or capillaries. Thirty minutes after application, target vessels and thesurrounding supporting tissue (e.g. skin) are removed. Biopsies arefixed in 10% paraformaldehyde, paraffin-embedded, and cut into 5-umsections on a microtome in transverse directions. Slides with mountedparaffin sections are deparaffinized and stained with hematoxylin andeosin (H&E) or silver enhancement staining. Using H&E staining and lightmicroscopy, one or several vessels, microvessels, and capillaries can beimaged. Scoring is performed for visible thermal damage of the vesselstructures. Additionally, vessel staining (e.g. CD31 stain) is performedto clearly identify vascular structures within tissue samples.

Example 7. Determination of Efficiency of Conversion of Light to ThermalEnergy

In one embodiment, a suspension of plasmonic nanoparticles(silica-coated nanoplates having a diameter of about 100-200 nm, asdescribed here) was prepared by formulating the plasmonic nanoparticlesin 20% propylene glycol in water to a concentration of about 1000 O.D.,and the ability of this suspension to convert laser light to thermalenergy was determined. Available commercial and research products, e.g.,stock solutions of carbon lotion (20-200 mg/ml carbon, TelsarSoftLight),Meladine spray (1 mg/ml melanin, Creative Technologies), Indocyaninegreen (5 mg/ml in water, Sigma Aldrich), and vehicle control (20%propylene glycol in water) were also tested. All solutions were diluted1:1 000 from their indicated stock solution concentration, loaded at 90μl per well into a 96-well plate, and baseline temperatures weremeasured by K thermocouple with micrometer (ExTech Instruments, WalthamMass.) and recorded. Solutions were then irradiated with repeated laserpulses at various wavelengths (e.g., 1064 nm, 810 nm, and 755 nm),fluence (e.g., 10, 20, and 30 J/cm2) and pulse sequence parameters(e.g., 30 ms and 55 ms). Following each sequential laser pulse, up to atotal of 8 pulses, solution temperatures were measured and recorded. Asshown in FIGS. 11A-11B, a series of plasmonic nanoparticle (PNP)formulations (labeled SL-001 and SL-002) exhibited ultra-high absorptioncompared to existing commercial and research chromophores. (FIGS. 11A,B) Rate of temperature increase over sequential laser pulses for PNPformulation SL-001 (FIG. 11A, closed circle), resonant at 1064 nm laserwavelength, upon irradiation with 1064 nm laser (A), and SL-002 (FIG.11B closed circle), resonant at 810 nm laser wavelength, uponirradiation with 810 nm laser (B). Control solutions are as follows:Carbon lotion (open triangle), Meladine spray (closed square),Indocyanine green (open diamond), and 20% propylene glycol (closedtriangle). All solutions were diluted 1:1000 from stock clinicalconcentration for laser irradiation and temperature measurements. For A,n=2 and error bars are s.d. of the mean.

Example 8. Quantitation of Nanoparticle Delivery into Target Tissues

In one embodiment, red fluorescent nanoparticles (Corpuscular Inc., ColdSpring, N.Y.) were contacted with isolated porcine skin explants asfollows. A 2.5 mg/ml solution of SiO₂, 200 nm diameter, 548 nm emissionparticles in 20% propylene glycol was pipetted onto the skin surface andmechanically massaged into the tissue explant. An ethanol wipe was usedto remove non-penetrating particles. As shown in FIGS. 12A-12B, theprovided formulations of nanoparticles (NPs) deeply and specificallypenetrate ex vivo porcine skin. FIG. 12A demonstrates representativesurvey fluorescence image of porcine skin, treated with red fluorescentNPs and histologically sectioned. Red (light contrast) NPs are imagedafter penetrating the hair follicle infundibulum (arrows) and deepfollicle, but not in the underlying dermis. FIG. 12B showsrepresentative confocal images show red NPs within superficial and deepfollicle (−870/−tm) at high and low magnification. Green (dark contrast)is tissue autofluorescence (488 nm emission). Scale bars as labeled 1 mm(A), 10 μm (B, left), 50 μm (B, right).

Further, formulations of nanoparticles (NPs) with silica coating deeplyand specifically penetrate in vivo human skin. A region of an upper armof a male human subject having skin Type 3 was treated with the rednanoparticles essentially as described above. Shown in FIGS. 13A and 13Bare representative confocal images of biopsies taken from the invivo-treated human skin, which were sectioned and immunostained for skinmarkers. Left-‘TH 2A R med’ sample shows red hair follicle fluorescenceafter red NP application with massage, ultrasound, and no pre-depilationwith waxing; Middle ‘TH 2C L’ sample shows red hair folliclefluorescence after red NP application with massage, ultration, andpre-depilation with waxing; Right—‘TH 1A Control’ shows background redautofluorescence of hair follicle. FIG. 13A is 3 color image where redis NPs, blue is collagen IV (staining basement membrane) and green isPGP 9.5 (staining nerve fiber). FIG. 13B shows red channel only in blackand white. Scale bars as labeled 100 μm.

Example 9. Treatment of Atrophic Scars with Needle Nose Tube Dispenser

In one embodiment, the following treatments were administered to theface of a patient using a ND:YAG laser and a composition containingsilica-coated silver plasmonic nanoplates delivered from a needle-nosetube dispenser. Silica-coated silver plasmonic nanoplates weresynthesized to have a resonant absorption peak at 1050 nm and formulatedin a cosmetic carrier containing water, propylene glycol, sodium dodecylsulfate, aristoflex AVC, and PE9010. Particle concentration was broughtto between 10¹² to 10¹³ particles per ml to achieve an optical densityof 100 O.D. at 1050 nm. The solution was dispensed from a needle nosetube dispenser into an ice pick acne scar and a box car chicken pox scarwith diameters of ˜1 mm and ˜3 mm respectively and a cosmetic sponge wasused to wipe away/soak up material that contacted the skin outside ofthe target region. Presence of the solution on and within the scars wasvisualized by its green hue. Electromagnetic radiation (light) wasadministered to the treatment areas after application of solution to theskin using an ND:YAG laser operating at a 5 ms pulse width, 7 mm spotsize, and a fluence of 15 J/cm². Two treatments were administered toeach scar at 1 week apart.

Treatment areas initially showed minimal pinkness of the skin, with nopurpura. After 24 hours the treated spots turned dark immediately aroundthe target region, showing increased pigmentation. Within 3-5 days oftreatment the darker spots scaled off and the skin returned to a normalappearance. After two weeks from the second treatment the diameter,depth, and prominence of both treated scars was noticeably reduced. (SeeFIG. 1) The series of images at the top of FIG. 14 shows an ice pickacne scar from baseline to 3 weeks post initial treatment. Selectivedelivery of photoactive material (silica-coated silver plasmonicnanoparticles) is visualized at baseline (Solution). The series ofimages at the bottom of FIG. 14 shows a box car chicken pox scar frombaseline to 3 weeks post initial treatment. Selective delivery ofphotoactive material (silica-coated silver plasmonic nanoparticles) isvisualized at baseline (Solution).

Example 10. Treatment of Atrophic Scars by Large Region Application andSelective Removal

In one embodiment, the following treatments were administered to theface of a patient using a 1064 nm ND:YAG laser and a compositioncontaining silica-coated silver plasmonic nanoplates delivered from atopical syringe. Silica-coated silver nanoplates were synthesized tohave a resonant absorption peak near 1050 nm and formulated in acosmetic carrier containing water, propylene glycol, sodium dodecylsulfate, aristoflex AVC, and PE9010. Particle concentration was broughtto between 10¹² to 10¹³ particles per ml to achieve an optical densityof 100 O.D. at 1050 nm. The solution was dispensed from a topicalsyringe to the cheeks of a patient with multiple atrophic acne scars todisperse the solution into each scar. A wet cloth was used to removephotoactive compound on the surface of the skin, but not localized inthe atrophic lesions. Presence of the solution on and within the scarswas visualized by its dark hue resulting in a speckled pattern on theface. Electromagnetic radiation (light) was administered to thetreatment areas after application of solution to the skin using a 1064nm ND:YAG laser operating at a 5 ms pulse width, 7 mm spot size, and afluence of 15 J/cm². Six treatments were administered at 2 weeks apart.

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. After 24 hours the treated spots turneddark immediately around the target region, showing increasedpigmentation. Within 3-5 days of each treatment the darker spots scaledoff and the skin returned to a normal appearance. After two weeks fromthe sixth treatment the diameter, depth, and prominence of atrophicscars was significantly reduced.

Example 11. Treatment of Atrophic Scars with Nanoshells

In one embodiment, the following treatments were administered to theface of a patient using a 755 nm Alexandrite laser and a compositioncontaining PEG-coated gold plasmonic nanoshells delivered from a topicalsyringe. PEG-coated gold plasmonic nanoshells were synthesized to have aresonant absorption peak near 750 nm and formulated in a cosmeticcarrier containing water, propylene glycol, aristoflex AVC, and PE9010.Particle concentration was brought to approximately 3×10¹¹ particles perml to achieve an optical density of 100 O.D. at 750 nm. The solution wasdispensed from a needle-nose applicator to individual atrophic acnescars on a patients phase and a sponge was used to wipe away/soak upmaterial in non-target regions. Presence of the solution on and withinthe scars was visualized by its dark hue resulting in a speckled patternon the face. Electromagnetic radiation (light) was administered to thetreatment areas after application of solution to the skin using a 755 nmAlexandrite laser operating at a 5 ms pulse width, 7 mm spot size, and afluence of 15 J/cm². Six treatments were administered at 2 weeks apart.

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. After 24 hours the treated spots turneddark immediately around the target region, showing increasedpigmentation. Within 3-5 days of each treatment the darker spots scaledoff and the skin returned to a normal appearance. After two weeks fromthe sixth treatment the diameter, depth, and prominence of atrophicscars was significantly reduced.

Example 12. Treatment of Pigmented Lesions/Freckles with SilverNanoplates

In one embodiment, the following treatments were administered to the armof a patient using a 755 nm alexandrite laser and a compositioncontaining silica-coated silver plasmonic nanoplates delivered from atopical syringe. Silica-coated silver plasmonic nanoplates weresynthesized to have a resonant absorption peak at 750 nm and formulatedin a cosmetic carrier containing water, propylene glycol, sodium dodecylsulfate, aristoflex AVC, and PE9010. Particle concentration was adjustedto between 10¹² to 10¹³ particles per ml to achieve an optical densityof 100 O.D. at 750 nm. The solution was dispensed from a needle noseapplicator to 3 individual sun spots/freckles on the arm of a patient.Presence of the solution on the freckles was visualized by its blue hueresulting in a speckled pattern on the arm. Electromagnetic radiation(light) was administered to the treatment areas after application ofsolution to the skin using an 755 nm Alexandrite laser operating at a0.5 ms, 5 ms, and 50 ms pulse width for each spot respectively, 7 mmspot size, and a fluence of 15 J/cm².

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. Pain and pinkness was the least in thearea treated with the 0.5 ms pulse width. After 24 hours 5 ms and 50 mstreated spots turned dark immediately around the target region, showingincreased pigmentation. Within 3-5 days of each treatment the darkerspots scaled off and the skin returned to a normal appearance. Withintwo weeks significant reduction in color/darkness of the pigmentedlesions/freckles was observed for spots treated with 5 ms and 50 mspulse widths. The color/darkness of the pigmented lesion treated with0.5 ms pulse was reduced, but not as prominently as the areas treatedwith 5 ms and 50 ms pulse widths.

Example 13. Treatment of Pigmented Lesions/Freckles with Nanorods

In one embodiment, the following treatments were administered to the armof a patient using a 755 nm alexandrite laser and a compositioncontaining PEG-coated gold plasmonic nanorods delivered from a topicalsyringe. PEG-coated gold plasmonic nanorods were synthesized to have aresonant absorption peak at 750 nm and formulated in a cosmetic carriercontaining water, propylene glycol, sodium dodecyl sulfate, aristoflexAVC, and PE9010. Particle concentration was adjusted to achieve anoptical density of 50 O.D. at 750 nm. The solution was dispensed from aneedle nose applicator to 3 individual sun spots/freckles on the arm ofa patient. Presence of the solution on the freckles was visualized byits dark hue resulting in a speckled pattern on the arm. Electromagneticradiation (light) was administered to the treatment areas afterapplication of solution to the skin using an 755 nm Alexandrite laseroperating at a 0.5 ms, 5 ms, and 50 ms pulse width for each spotrespectively, 7 mm spot size, and a fluence of 15 J/cm².

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. Pain and pinkness was least in the areatreated with the 0.5 ms pulse width. After 24 hours 5 ms and 50 mstreated spots turned dark immediately around the target region, showingincreased pigmentation. Within 3-5 days of each treatment the darkerspots scaled off and the skin returned to a normal appearance. Withintwo weeks significant reduction in color/darkness of the pigmentedlesions/freckles was observed for spots treated with 5 ms and 50 mspulse widths. The color/darkness of the pigmented lesion treated with0.5 ms pulse was reduced, but not as prominently as the areas treatedwith 5 ms and 50 ms pulse widths.

Example 14. Treatment of Pigmented Lesions/Freckles with CarbonNanoparticles

In one embodiment, the following treatments were administered to the armof a patient using a 1064n ND:YAG laser and a composition containingcarbon nanoparticles. Carbon nanoparticles were coated with polyvinylpyrrolidone and formulated in a cosmetic carrier containing water,propylene glycol, sodium dodecyl sulfate, aristoflex AVC, and PE9010.Particle concentration was brought to 1-5 mg per ml to achieve anoptical density of 100 O.D. at 1050 nm. The solution was dispensed froma needle nose applicator to 3 individual sun spots/freckles on the armof a patient. Presence of the solution on the freckles was visualized byits black hue resulting in a speckled pattern on the arm.Electromagnetic radiation (light) was administered to the treatmentareas after application of solution to the skin using an 1064 nm ND:YAGlaser operating at a 0.5 ms, 5 ms, and 50 ms pulse width for each spotrespectively, 7 mm spot size, and a fluence of 15 J/cm².

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. Pain and pinkness was least in the areatreated with the 0.5 ms pulse width. After 24 hours 5 ms and 50 mstreated spots turned dark immediately around the target region, showingincreased pigmentation. Within 3-5 days of each treatment the darkerspots scaled off and the skin returned to a normal appearance. Withintwo weeks significant reduction in color/darkness of the pigmentedlesions/freckles was observed for spots treated with 5 ms and 50 mspulse widths. The color/darkness of the pigmented lesion treated with0.5 ms pulse was reduced, but not as prominently as the areas treatedwith 5 ms and 50 ms pulse widths.

Example 15. Treatment of Striae

The following treatments were administered to the abdomen of a patientusing a 1064 nm ND:YAG laser and a composition containing silica-coatedsilver plasmonic nanoplates delivered from a topical syringe.Silica-coated silver plasmonic nanoplates were synthesized to have aresonant absorption peak at 1050 nm and formulated in a cosmetic carriercontaining water, propylene glycol, sodium dodecyl sulfate, aristoflexAVC, and PE9010. Particle concentration was brought to between 10¹² to10¹³ particles per ml to achieve an optical density of 100 O.D. at 1050nm. The solution was dispensed from a needle nose applicator alongmultiple lines of striae on a patient. Presence of the solution on thestriae was visualized by its green hue. Electromagnetic radiation(light) was administered to the treatment areas after application ofsolution to the skin using an 1064 nm ND:YAG laser operating at a 5 mspulse width, 7 mm spot size, and a fluence of 15 J/cm². Three treatmentswere administered at 2 weeks apart.

Treatment areas initially showed minimal pinkness of the skin, with nopurpura after each treatment. After 24 hours the treated areas turneddark immediately around the target region, showing increasedpigmentation. Within 3-5 days of each treatment the darker spots scaledoff and the skin returned to a normal appearance. Appearance of striaewas noticeably reduced two weeks after the third treatment.

Example 16. Composition Delivery Devices

In an experiment, an embodiment of a composition 100 of nanoparticleswas distributed to various target tissue depths with various embodimentsof delivery devices 200. In the experiment, an animal skin model (pigear) comprising hair follicles and sebaceous glands in an epidermis wasused to model skin treatment with a composition 100. In variousembodiments, ultrasound and/or other sonic forces, mechanicalvibrations, hair shaft manipulation (including pulling), physical force,thermal manipulation, and other treatments are utilized to improve entryof a composition 100 of light-absorbing nanoparticles into hairfollicles and/or sebaceous glands. Mechanical vibration massage improvesfollicular penetration through hair shaft ‘pumping’ mechanisms, whileultrasound enhances transdermal drug delivery through temporarydisruption of the skin's lipid bilayer, bubble formation, liquidmicrostreaming, jetting, streaming, heating, and/or cavitation. Acomposition 100 of plasmonic nanoparticles was contacted with the skinmodel to produce a semi-quantitative assay for measuring and optimizingdelivery efficacy. Through experimentation, evaluation of formulationsof the composition 100 in conjunction with testing of individual and/orcombinations of embodiments of delivery devices 200.

FIG. 15 illustrates an embodiment of a delivery device 200 distributinga composition 100 from a skin surface to a target skin depth. In theillustrated FIG. 15, the target skin comprises a hair follicle and/or asebaceous gland. In one embodiment, FIG. 15 is a schematic side view ofa composition being distributed from a skin surface to a target in thetissue with a delivery device according to an embodiment of theinvention (left side). The formulation (100) is applied to the skinfollowed by the delivery device (200), e.g., massage or ultrasoundtreatment. (right side) After wiping away excess formulation from thesurface of the skin, composition (100) is localized only within thestructures of the hair follicle (e.g., infundibulum, lumen, sebaceousgland) and no longer on the surface of the epidermis. In one embodiment,the follicle is a plugged follicle. Three embodiments of deliverydevices 200 were used. For each of the delivery devices 200, acomposition 100 was applied to the surface of the skin model. Thedelivery device 200 was activated to move the composition 100 from theskin surface to the target tissue depth. As illustrated at theembodiment at FIG. 16, the skin model was sectioned into 40 micronslices, which were then mounted and imaged using a high resolution colorscanner. In one embodiment, FIG. 16 includes: (A) an illustration of theskin after targeted delivery of the composition (100). (B) Using commonhistological protocols, a smaller section of the skin that was treatedis fixed and sectioned into horizontal slices between 10-60 micronsthick. Areas where the formulation (100) was delivered can be observedwithin the individual hair follicles contained within these slices. (C)The individual tissue sections are mounted for imaging. (D) Each tissuesection is imaged using a high resolution optical scanner. Theformulation (100) can be observed (blue color) without staining. (E)Analysis of percent delivery is calculated as the ratio of theformulation (blue color) to the total area of the section and expressedas a percentage.

The mounted section of skin is imaged. In one embodiment, thecomposition 100 is blue. The images are analyzed measuring the totalblue area as a ratio to the total section area. Delivery measurementsare quantified as the ratio of blue area (composition 100) over thetotal area. In various embodiments, the composition 100 can be anycolor.

FIG. 17 is a table summarizing experimental results using threeembodiments of delivery devices 200 to deliver a composition 100 tovarious depths in tissue according to various embodiments. The vibraderm(210) is an adjustable vibration device applying a nominal 80 Hzlongitudinal vibration. The Acoustic Horn/Sonotrode (220) is a 30-40 kHzultrasound device employing an acoustic horn or sonotrode to apply theultrasonic energy into the composition. Ultrasound from an acoustic hornor sonotrode is focused in the volume immediately surrounding the hornor sonotrode and therefore produces cavitation mostly in the formulationand at the surface of the skin. The flat transducer (230) is a 30-40 kHzultrasound device employing a flat head to deliver the ultrasonic energyinto the formulation. Ultrasound from a flat transducer is “unfocused”in that it produces cavitation at various distances away from theultrasound head. The first delivery device 210 is a Vibraderm with 80 Hzlongitudinal vibration. The second delivery device 220 is a 30 kHz-40kHz low frequency ultrasound device with a sonotrode that operates at afrequency about 32.4 kHz pulsed ultrasound with surface localized energyconfigured to induce cavitation (IMPACT, Alma Lasers). The thirddelivery device 230 is a low frequency ultrasound device that operatesat a frequency of 40 kHz non-pulsed ultrasound configured to inducecavitation from an unfocused transducer (GS8.0). The first deliverydevice 210 uses a mechanical mixing mechanism resulting in a 4% of totalskin cross-section ratio measurement of composition 100 at a depth of500 micrometers in porcine ear skin, with a maximum delivery depth ofabout 1000 microns in porcine ear skin. The second delivery device 220uses a cavitation mechanism resulting in a 12% of total skincross-section ratio measurement of composition 100 at a depth of 500micrometers in porcine ear skin, with a maximum delivery depth of about1500 microns in porcine ear skin. The third delivery device 230 uses acavitation mechanism with an unfocused transducer resulting in avariable 4-12% of total skin cross-section ratio measurement ofcomposition 100 at a depth of 500 micrometers in porcine ear skin, witha maximum delivery depth of about 1000 microns in porcine ear skin.

FIG. 18 illustrates images of delivery of composition 100 to skin models(pig ears) using three delivery techniques: low frequency ultrasound (at40 kHz for 1 minute), massage by hand (for 1 minute), and throughincubation on the skin surface (placement of the composition 100 on theskin surface for 5 minutes). The top panel contains 5× brightfieldimages looking down at the surface of intact skin models immediatelyfollowing the delivery of an embodiment of the composition. In theleftmost image of the top panel, low frequency ultrasound, theformulation (100) is observed to be localized within several hairfollicles. In the middle and rightmost images of the top panel, massageby hand and incubate on skin respectively, no delivery is observed. Thelower panel contains 20× brightfield images of vertical cross sectionsof the skin. The cross sections are cut to expose an individual hairfollicle. In the leftmost image of the bottom panel, low frequencyultrasound, the formulation (100) is observed clearly localized withinthe infundibulum, sebaceous gland, and the lumen of the hair follicle.In the middle and rightmost images of the bottom panel, massage by handand incubate on skin respectively, no delivery is limited to theuppermost region of the infundibulum of the hair follicles. Asillustrated in the images, the composition 100 penetrates the deepestwith the low frequency ultrasound device, with the composition 100delivered to the infundibulum (INF), sebaceous gland (SG) and the hairfollicle lumen (HFL). As illustrated in the images, the composition 100penetrates to the infundibulum (INF) with hand massage. As illustratedin the images, the composition 100 penetrates near the skin surface.

FIG. 19 is a graph plotting data from the experiment, showing the totalpercent positive area on the y-axis, and depth in microns on the x-axis.Plotted are a negative control (incubation on the skin surface without adelivery device 200), the first delivery device 210 (Vibraderm), and acombination of both the first delivery device 210 (Vibraderm) and thesecond delivery device 220 (30 kHz-40 kHz ultrasound device with asonotrode). FIG. 20 is a graph plotting data from the experiment,showing the total percent positive area on the y-axis, and depth inmicrons on the x-axis. Plotted are a negative control (incubation on theskin surface without a delivery device 200), the first delivery device210 (Vibraderm), a combination of both the first delivery device 210(Vibraderm) and the third delivery device 230 (36 kHz non-pulsedultrasound), and a combination of both the first delivery device 210(Vibraderm) and the second delivery device 220 (30 kHz-40 kHz ultrasounddevice with a sonotrode).

FIG. 21 includes images from unstained horizontal cross-sections at 720microns in depth below the skin surface, illustrating the distributionof composition 100 from the experiment. The images include distributionof the composition 100 for a negative control (incubation on the skinsurface without a delivery device 200), the first delivery device 210(Vibraderm), a combination of both the first delivery device 210(Vibraderm) and the third delivery device 230 (36 kHz non-pulsedultrasound), and a combination of both the first delivery device 210(Vibraderm) and the second delivery device 220 (30 kHz-40 kHz ultrasounddevice with a sonotrode). FIG. 21 is illustrative of images taken oftissue sections at approximately 720 microns deep of compositiondelivery within tissue samples using various embodiments of deliverydevices. For the delivery device embodiment of a vibration device(Vibraderm) plus acoustic horn/sonotrode ultrasound energy, there is amarkedly increase in the total amount of composition delivered (100) andnumber of follicles that the composition is delivered to over the otherdelivery embodiments alone. At levels of approximately 720 microns,there is also a significant increase in delivery of the composition withvibration device (Vibraderm) plus 30-40 kHz ultrasound (flat transducer)over the vibration device alone.

FIG. 22 includes images from unstained horizontal cross-sections at 1060microns in depth below the skin surface, illustrating the distributionof composition 100 from the experiment. The images include distributionof the composition 100 for a negative control (incubation on the skinsurface without a delivery device 200), the first delivery device 210(Vibraderm), a combination of both the first delivery device 210(Vibraderm) and the third delivery device 230 (rey), and a combinationof both the first delivery device 210 (Vibraderm) and the seconddelivery device 220 (30 kHz-40 kHz ultrasound device with a sonotrode).FIG. 22 is illustrative of images taken of tissue sections atapproximately 1060 microns deep of composition delivery within tissuesamples using various embodiments of delivery devices. For the deliverydevice embodiment of a vibration device plus acoustic horn/sonotrodeultrasound energy, there is a markedly increase in the total amount ofcomposition delivered (100) and number of follicles that the compositionis delivered to over the other delivery embodiments alone. At this depthof approximately 1060 microns, the amount delivery of the composition bythe other delivery device embodiments has significantly decreased.

Unless otherwise indicated, all numbers expressing quantities ofingredients, reaction conditions, and so forth used in the specificationand claims are to be understood as being modified in all instances bythe term “about.” Accordingly, unless indicated to the contrary, thenumerical parameters set forth in the following specification andattached claims are approximations that may vary depending upon thedesired properties sought to be obtained by the present invention.

Other embodiments of the invention will be apparent to those skilled inthe art from consideration of the specification and practice of theinvention disclosed herein. It is intended that the specification andexamples be considered as disclosing certain embodiments of theinvention only, with a true scope and spirit of the invention beingindicated by the following claims.

As will be understood by the skilled artisan, the subject matterdescribed herein may be embodied in other specific forms withoutdeparting from the spirit or essential characteristics thereof. Theforegoing embodiments are therefore to be considered in all respectsillustrative rather than limiting on the invention described herein.While the invention is susceptible to various modifications, andalternative forms, specific examples thereof have been shown in thedrawings and are herein described in detail. It should be understood,however, that the invention is not to be limited to the particular formsor methods disclosed, but to the contrary, the invention is to cover allmodifications, equivalents, and alternatives falling within the spiritand scope of the various embodiments described and the appended claims.Any methods disclosed herein need not be performed in the order recited.The methods disclosed herein include certain actions taken by apractitioner; however, they can also include any third-party instructionof those actions, either expressly or by implication. For example,actions such as “identifying a target region of skin tissue” include“instructing the identification of a target region of skin tissue.” Theranges disclosed herein also encompass any and all overlap, sub-ranges,and combinations thereof. Language such as “up to,” “at least,” “greaterthan,” “less than,” “between,” and the like includes the number recited.Numbers preceded by a term such as “about” or “approximately” or“substantially” include the recited numbers. For example, “about 3 mm”includes “3 mm.” The terms “approximately”, “about”, and “substantially”as used herein represent an amount or characteristic close to the statedamount or characteristic that still performs a desired function orachieves a desired result. For example, the terms “approximately”,“about”, and “substantially” may refer to an amount that is within lessthan 10% of, within less than 5% of, within less than 1% of, within lessthan 0.1% of, and within less than 0.01% of the stated amount orcharacteristic.

What is claimed:
 1. A method of delivering a composition to a targettissue under a skin surface with a delivery device, comprising: applyinga composition to a skin surface, distributing the composition from theskin surface to a target tissue under the skin surface with a deliverydevice, wherein the delivery device is an ultrasound device; whereinsaid composition comprises a plurality of unassembled plasmonicnanoparticles, wherein the unassembled plasmonic nanoparticles comprisea conductive metal portion, wherein the conductive metal portioncomprises at least one of gold or silver, wherein the unassembledplasmonic nanoparticles have a size in a range of 10 nm to 300 nm,wherein the unassembled plasmonic nanoparticles comprise a coating thatcoats the conductive metal portion, wherein said coating facilitatesselective removal from the skin surface; wherein the coating comprisesat least one of silica or polyethylene glycol (PEG), selectivelyremoving the composition from the skin surface, while leaving thecomposition localized within the sebaceous gland; and irradiating thecomposition with an infrared light source thereby inducing a pluralityof surface plasmons in said unassembled plasmonic nanoparticles, whereininducing the plurality of surface plasmons generates localized heat inthe target tissue.
 2. The method of claim 1, wherein the delivery deviceis a sonic device.
 3. The method of claim 1, further comprisingpre-treating the skin surface prior to irradiating the composition,wherein pre-treating the skin surface comprises hair removal.
 4. Themethod of claim 1, wherein the unassembled plasmonic nanoparticles havea concentration of 10⁹ to 10¹³ particles per ml of the composition. 5.The method of claim 1, wherein the unassembled plasmonic nanoparticlescomprise a solid, conducting silver core and a silica coating.
 6. Themethod of claim 1, wherein the conductive metal portion is a silvernanoplate, and wherein the coating is less conductive than theconductive metal portion.
 7. The method of claim 1, wherein theconductive metal portion is a nanoplate, and wherein the nanoplate has apeak absorption wavelength in a range of 750 nm to 1200 nm.
 8. Themethod of claim 1, wherein the coating comprises silica, wherein thetarget tissue comprises at least one of a sebocyte and sebum.
 9. Amethod of delivering a composition to a sebaceous gland and performingablation, comprising: topically applying a solution of unassembledplasmonic nanoparticles to a skin surface, targeting a sebaceous glandby redistributing the solution of unassembled plasmonic nanoparticlesfrom the skin surface to the sebaceous gland with a delivery device;wherein the delivery device is a sonic mechanical vibration device;wherein the unassembled plasmonic nanoparticles have a dimension in arange of 10 nm to 300 nm, wherein the unassembled plasmonicnanoparticles comprise a conductive metal portion, wherein theconductive metal portion comprises at least one of gold or silver,wherein the unassembled plasmonic nanoparticles comprise a coating thatcoats the conductive metal portion, wherein said coating facilitatesselective removal from the skin surface; wherein the coating comprisesat least one of silica or polyethylene glycol (PEG), selectivelyremoving the solution from the skin surface, while leaving the solutionlocalized within the sebaceous gland; and irradiating the solution ofunassembled plasmonic nanoparticles with an energy wavelength in a rangeof 750 nm to 1200 nm to induce a plurality of surface plasmons in saidunassembled plasmonic nanoparticles to cause ablation in the sebaceousgland, thereby treating acne at said sebaceous gland.
 10. The method ofclaim 9, wherein the redistributing the solution with the sonicmechanical vibration device comprises bubble formation or liquidmicrostreaming.
 11. The method of claim 9, further comprisingpre-treating the skin surface to increase delivery of the unassembledplasmonic nanoparticles to the sebaceous gland with at least one of thegroup consisting of shaving, waxing, peeling, cyanoacrylate surfacepeeling, a calcium thioglycolate treatment, a surface exfoliation, amechanical exfoliation, a salt glow, a microdermabrasion, a chemicalexfoliation, a chemical exfoliation with an enzyme, a chemicalexfoliation with alphahydroxy acid, and a chemical exfoliation withbetahydroxy acid.
 12. The method of claim 9, wherein the concentrationof the unassembled plasmonic nanoparticles is 10¹¹ to 10¹³ particles perml of the solution, wherein the coating is less conductive than theconductive metal portion.
 13. The method of claim 9, wherein theunassembled plasmonic nanoparticles have an optical density of 10 O.D.to 5,000 O.D. within an infrared light range.
 14. The method of claim 9,wherein the coating is semiconductive, wherein the conductive metalportion is inside the coating, and wherein the coating is lessconductive than the conductive metal portion.
 15. The method of claim 9,wherein the conductive metal portion is a nanoplate, and wherein thecoating is less conductive than the conductive metal portion.
 16. Amethod of delivering a composition of unassembled plasmonicnanoparticles to a pilosebaceous unit and performing ablation,comprising: applying a solution of unassembled plasmonic nanoparticlesto a skin surface, distributing the solution of unassembled plasmonicnanoparticles with a mechanical vibration device from the skin surfaceto a pilosebaceous unit thereby targeting the pilosebaceous unit,wherein the mechanical vibration device comprises at least one of thegroup consisting of a sonic force device, a high pressure air flowdevice, a high pressure liquid flow device, a vacuum device, and adermabrasion device, wherein the pilosebaceous unit comprises one ormore structures consisting of: a hair shaft, a hair follicle, asebaceous gland, and a hair follicle infundibulum; wherein theunassembled plasmonic nanoparticles comprise a conductive metal portion,wherein the conductive metal portion comprises at least one of gold orsilver; wherein the unassembled plasmonic nanoparticles have a peakabsorption wavelength of between 750 nm and 1200 nm, wherein theunassembled plasmonic nanoparticles comprise a coating that coats theconductive metal portion, wherein the coating comprises at least one ofsilica or polyethylene glycol (PEG), selectively removing the solutionfrom the skin surface while leaving the solution localized within theportion of the sebaceous gland, and irradiating the solution with anenergy to induce said unassembled plasmonic nanoparticles to generatelocalized ablation, wherein said ablation comprises thermal damage insaid sebaceous gland.
 17. The method of claim 16, further comprising:pre-treating the skin surface to increase delivery of the unassembledplasmonic nanoparticles to the pilosebaceous unit with at least one ofthe group consisting of shaving, waxing, peeling, cyanoacrylate surfacepeeling, a calcium thioglycolate treatment, a surface exfoliation, amechanical exfoliation, a salt glow, a microdermabrasion, a chemicalexfoliation, a chemical exfoliation with an enzyme, a chemicalexfoliation with alphahydroxy acid, and a chemical exfoliation withbetahydroxy acid; wherein irradiating the solution of unassembledplasmonic nanoparticles comprises exposing the solution of unassembledplasmonic nanoparticles to the energy at a wavelength of between 750 nmand 1200 nm to induce a plurality of surface plasmons in saidunassembled plasmonic nanoparticles, thereby treating acne at saidsebaceous gland.
 18. The method of claim 16, further comprising: whereinthe mechanical vibration device is a sonic force device; whereinirradiating the solution of unassembled plasmonic nanoparticles with theenergy comprises an infrared light source wavelength of between 750 nmand 1200 nm to induce a plurality of surface plasmons in saidunassembled plasmonic nanoparticles, thereby treating acne at saidsebaceous gland.
 19. The method of claim 16, wherein the unassembledplasmonic nanoparticles are nanoplates, wherein the unassembledplasmonic nanoparticles have an optical density of 10 O.D. to 5,000 O.D.within an infrared light range and the concentration is 10⁹ to 10¹³particles per ml of the solution; and wherein irradiating the solutionof unassembled plasmonic nanoparticles with the energy comprises aninfrared wavelength of between 750 nm and 1200 nm to induce a pluralityof surface plasmons in said unassembled plasmonic nanoparticles, therebyheating said pilosebaceous unit.
 20. The method of claim 16, whereinselectively removing the composition from the skin surface comprisesusing water or alcohol to remove the composition from the skin surfacewhile leaving the composition localized within the pilosebaceous unit.